New isothiazoloquinolones and related compounds as anti-infective agents

ABSTRACT

The invention provides certain compounds and salts of Formula I and Formula II: 
     
       
         
         
             
             
         
       
     
     which possess antimicrobial activity. The invention also provides novel synthetic intermediates useful in making compounds of Formula I and Formula II. The variables A 1 , R 2 , R 3 , R 5 , R 6 , R 7 , A 8  and R 9  are defined herein. 
     Certain compounds of Formula I and Formula II disclosed herein are potent and selective inhibitors of bacterial DNA synthesis and bacterial replication. The invention also provides antimicrobial compositions, including pharmaceutical compositions, containing one or more compounds of Formula I or Formula II and one or more carriers, excipients, or diluents. Such compositions may contain a compound of Formula I or Formula II as the only active agent or may contain a combination of a compound of Formula I or Formula II and one or more other active agents. The invention also provides methods for treating microbial infections in animals.

CROSS REFERENCE TO RELATED APPLICATION

This application claims priority from U.S. Provisional Application No.60/653,434 filed Feb. 16, 2005, which is hereby incorporated byreference in its entirety.

FIELD OF THE INVENTION

The present invention provides isothiazolo[5,4-b]quinolones and relatedcompounds, which possess antimicrobial activity. Certain compoundsprovided herein possess potent antibacterial, antiprotozoal, orantifungal activity. Particular compounds provided herein are alsopotent and/or selective inhibitors of prokaryotic DNA synthesis andprokaryotic reproduction. The invention provides anti-microbialcompositions, including pharmaceutical compositions, containing one ormore carrier, diluents, or excipients. The invention providespharmaceutical compositions containing an isothiazolo[5,4-b]quinoline orrelated compound as the only active agent or containing anisothiazolo[5,4-b]quinoline or related compound in combination with oneor more other active agent, such as one or more other antimicrobial orantifungal agent. The invention provides methods for treating orpreventing microbial infections in eukaryotes, preferably animals, byadministering an effective amount of a isothiazolo[5,4-b]quinoline orrelated compound to a eukaryote suffering from or susceptible tomicrobial infection. The invention also provides methods of inhibitingmicrobial growth and survival by applying an effective amount of aisothiazolo[5,4-b]quinoline or related compound.

The invention also provides novel intermediates useful for the for thesynthesis of isothiazolo[5,4-b]quinolones and related compounds. Theinvention also provides methods of synthesis forisothiazolo[5,4-b]quinolones and related compounds.

BACKGROUND OF THE INVENTION

Antimicrobial compounds are compounds capable of destroying orsuppressing the growth or reproduction of microorganisms, such asbacteria, protozoa, mycoplasma, yeast, and fungi. The mechanisms bywhich antimicrobial compounds act vary. However, they are generallybelieved to function in one or more of the following ways: by inhibitingcell wall synthesis or repair; by altering cell wall permeability; byinhibiting protein synthesis; or by inhibiting synthesis of nucleicacids. For example, beta-lactam antibacterials inhibit the essentialpenicillin binding proteins (PBPs) in bacteria, which are responsiblefor cell wall synthesis. Quinolones act, at least in part, by inhibitingsynthesis of DNA, thus preventing the cell from replicating.

Many attempts to produce improved antimicrobials yield equivocalresults. Indeed, few antimicrobials are produced that are trulyclinically acceptable in terms of their spectrum of antimicrobialactivity, avoidance of microbial resistance, and pharmacology. Thusthere is a continuing need for broad-spectrum antimicrobials, and aparticular need for antimicrobials effective against resistant microbes.

Pathogenic bacteria are known to acquire resistance via several distinctmechanisms including inactivation of the antibiotic by bacterial enzymes(e.g., beta-lactamases that hydrolyze penicillin and cephalosporins);removal of the antibiotic using efflux pumps; modification of the targetof the antibiotic via mutation and genetic recombination (e.g.,penicillin-resistance in Neiserria gonorrhea); and acquisition of areadily transferable gene from an external source to create a resistanttarget (e.g., methicillin-resistance in Staphylococcus aureus). Thereare certain Gram-positive pathogens, such as vancomycin-resistantEnterococcus faecium, which are resistant to virtually all commerciallyavailable antibiotics.

Resistant organisms of particular note include methicillin-resistant andvancomycin- resistant Staphylococcus aureus, penicillin-resistantStreptococcus pneumoniae, vancomycin-resistant enterococci,fluoroquinolone-resistant E. coli, cephalosporin-resistant aerobicgram-negative rods and imipenem- resistant Pseudomonas aeruginosa. Theseorganisms are significant causes of nosocomial infections and areclearly associated with increasing morbidity and mortality. Theincreasing numbers of elderly and immunocompromised patients areparticularly at risk for infection with these pathogens. Therefore,there is a large unmet medical need for the development of newantimicrobial agents.

SUMMARY OF THE INVENTION

The invention provides compounds of Formula I and Formula II (shownbelow) and includes isothiazolo[5,4-b]quinolines and related compounds,which possess antimicrobial activity. The invention provides compoundsof Formula I and Formula II that possess potent and/or selectiveantibacterial, antiprotozoal, or antifungal activity. The invention alsoprovides anti-bacterial compositions containing one or more compounds ofFormula I or Formula II, or a salt, solvate, or acylated prodrug of sucha compound, and one or more carriers, excipients, or diluents.

The invention further comprises methods of treating and preventingmicrobial infections, particularly bacterial and protozoal infections byadministering and effective amount of a compound of Formula I or FormulaII to a eukaryote suffering from or susceptible to a microbialinfection. These microbial infections include bacterial infections, forexample E. coli infections, Staphylococcus infections, Salmonellainfections and protozoal infections, for example Chlamydia infections.The invention is particularly includes methods of preventing or treatingmicrobial infections in mammals, including humans, but also encompassesmethods of preventing or treating microbial infections in other animals,including fish, birds, reptiles, and amphibians.

Methods of treatment include administering a compound of Formula I orFormula II alone as the single active agent or administering a compoundof Formula I in combination with one or more other therapeutic agent,such as an antibacterial, an antifungal, an antiviral, an interferon, anefflux-pump inhibitor, a beta-lactamase inhibitor, or another compoundof Formula I or Formula II.

The invention also provides methods of inhibiting microbial growth andsurvival by applying an effective amount of anisothiazolo[5,4-b]quinoline or related compound. The invention includes,for example, methods of inhibiting microbial growth and survival onmedical instruments or on surfaces used for food preparation by applyinga composition containing a compound of Formula I or Formula II.

Thus, the invention include compounds of Formula I and Formula II

A compound of Formula I

and the pharmaceutically acceptable salts thereof, wherein:

A₁ is S, O, SO, or SO₂.

R₂ is hydrogen.

Or, R₂ is C₁-C₈alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl,C₃-C₇cycloalkyl(C₀-C₄carbohydryl), C₄-C₇cycloalkenyl(C₀-C₄carbohydryl),aryl(C₀-C₄carbohydryl), C₂-C₆heterocycloalkyl(C₀-C₄carbohydryl) each ofwhich is substituted with 0 to 5 substituents independently chosen fromhalogen, hydroxy, amino, cyano, nitro, C₁-C₄alkyl, C₁-C₄alkoxy,C₁-C₂haloalkyl, C₁-C₂haloalkoxy, mono- and di-C₁-C₄alkylamino,C₂-C₄alkanoyl, C₁-C₄alkylthio, ═NOR₁₀, ═NR₁₀, —O(C═O)R₁₀, —(C═O)NR₁₀R₁₁,—O(C═O)NR₁₀R₁₁, —(C═O)OR₁₀, —(C═O)NR₁₀OR₁₁, —NR₁₀(C═O)R₁₁,—NR₁₀(C═O)OR₁₁, —NR₁₀(C═O)NR₁₁R₁₂, —NR₁₀(C═S)NR₁₁R₁₂, —NR₁₀NR₁₁R₁₂,—SO₃R₁₀, —(S═O)OR₁₀, —SO₂R₁₃, —SO₂NR₁₀R₁₁, and —NR₁₀SO₂R₁₃; where R₁₀,R₁₁, and R₁₂ are independently hydrogen, C₁-C₄alkyl, or aryl, and R₁₃ isC₁-C₄alkyl or aryl.

R₃ is hydrogen, C₁-C₆alkyl, C₁-C₆alkanoyl, mono- ordi-C₁-C₆alkylcarbamate, or C₁-C₆alkylsulfonate; each of which issubstituted with 0 to 3 substituents independently chosen from halogen,hydroxy, amino, cyano, nitro, C₁-C₄alkoxy, mono- and di-C₁-C₄alkylamino,C₁-C₂haloalkyl, and C₁-C₂haloalkoxy.

R₅ is hydrogen, halogen, hydroxy, amino, cyano, nitro, or —NHNH₂.

Or, R₅ is C₁-C₄alkyl, C₁-C₄alkoxy, mono- or di-(C₁-C₄)alkylamino, mono-,di- or tri-C₁-C₄ alkylhydrazinyl, C₂-C₄alkanoyl, C₁-C₄alkylester,C₁-C₂haloalkyl, or C₁-C₂haloalkoxy; each of which is substituted with 0to 3 substituents independently chosen from hydroxy, amino, halogen,oxo, C₁-C₄alkoxy, C₁-C₂haloalkyl, C₁-C₂haloalkoxy, and mono- anddi-C₁-C₄alkylamino.

R₆ is hydrogen, halogen, hydroxy, amino, cyano, C₁-C₄alkyl, C₁-C₄alkoxy,mono- or di-(C₁-C₄)alkylamino, —SO₃R₁₀, —SO₂R₁₀, or —SO₂NR₁₀R₁₁; whereR₁₀ and R₁₁ carry the definitions set forth above.

R₇ is bromo, iodo, —O(SO₂)CF₃, or —N₂BF₄, or R₇ is XR_(A).

Where, X is absent, —CH₂—CH₂—, —CH═CH—, —(C═O)—, —(C═O)NH—, or —C≡C—.

R_(A) is C₃-C₆alkyl, C₄-C₇cycloalkyl, C₄-C₇cycloalkenyl, a 7-10 memberedbicyclic saturated, partially unsaturated, or aromatic carbocyclicgroup, a 5-6 membered saturated, partially unsaturated, or aromaticheterocylic group bound via a carbon atom when X is absent or —CH₂—CH₂—,or bound via a carbon or nitrogen atom when X is —CH═CH— or —C≡C— or aR_(A) is a 7-10 membered bicyclic saturated, partially unsaturated, oraromatic heterocylic group bound via a carbon atom when X is absent or—CH₂—CH₂—, or bound via a carbon or nitrogen atom when X is —CH═CH— or—C≡C—; each of which R_(A) is substituted with 0 to 5 substituentsindependently chosen from (i), (ii), and (iii).

Or, R₇ is XR_(B), where R_(B) is phenyl substituted with 1 to 5substituents independently chosen from (i), (ii), and (iii).

Or, R₇ is XR_(C), where R_(C) is cyclopropyl with 0 to 5 substituentsindependently chosen from (i), (ii), and (iii), with the proviso thatR_(C) is not substituted with amino, or mono- or di-(C₁-C₄)alkylamino.

Or, R₇ is XR_(D) where R_(D) is phenyl fused to a 5- or 6-memberedheterocycloalkyl ring containing 1 or 2 nitrogen or oxygen atoms, whereR_(D) is substituted with 0 to 3 substitutents chosen from (i), (ii),and (iii).

Where,

-   -   (i) is chosen from halogen, hydroxy, amino, cyano, and nitro,    -   (ii) is chosen from C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl,        C₁-C₆alkoxy(C₀-C₄alkyl), mono- and di-(C₁-C₄)alkylamino,        C₁-C₂haloalkyl, C₁-C₂haloalkoxy,        C₃-C₇cycloalkyl(C₀-C₄carbohydryl),        C₃-C₇cycloalkyl(C₀-C₄carbohydryl-O—),        C₄-C₇cycloalkenyl(C₀-C₄carbohydryl), aryl(C₀-C₆carbohydryl),        aryl(C₁-C₄alkoxy), C₂-C₆heterocycloalkyl(C₀-C₄carbohydryl),        heteroaryl(C₀-C₆carbohydryl), C₁-C₆alkylthio, ═NOR₁₀, ═NR₁₀,        —(C₀-C₄alkyl)(C═O)R₁₀, —(C₀-C₄alkyl)O(C═O)R₁₀,        —(C₀-C₄alkyl)(C═O)NR₁₀R₁₁, —(C₀-C₄alkyl)O(C═O)NR₁₀R₁₁,        —(C₀-C₄alkyl)(C═O)OR₁₀, —(C₀-C₄alkyl)NR₁₀(C═O)R₁₁,        —(C₀-C₄alkyl)NR₁₀(C═O)OR₁₁, —(C₀-C₄alkyl)NR₁₀(C═O)NR₁₁R₁₂,        —(C₀-C₄alkyl)NR₁₀(C═O)(C₁-C₄alkyl)NR₁₁(C═O)O—R₁₂,        —(C₀-C₄alkyl)NR₁₀(C═S)NR₁₁R₁₂, —(C₀-C₄alkyl)NR₁₀NR₁₁R₁₂,        —(C₀-C₄alkyl)N═NR₁₃, —(C₀-C₄alkyl)SO₃R₁₀,        —(C₀-C₄alkyl)(S═O)OR₁₀, —(C₀-C₄alkyl)SO₂R₁₃,        —(C₀-C₄alkyl)SO₂NR₁₀R₁₁, and —(C₀-C₄alkyl)NR₁₀SO₂R₁₃; and    -   (iii) is chosen from —OR_(D), —(C═O)R_(D), —SO₂R_(D), —SO₃R_(D),        —NR₁₀SO₂R_(D), where R_(D) is C₁-C₄alkyl,        C₃-C₁cycloalkyl(C₀-C₂alkyl), C₂-C₆heterocycloalkyl(C₀-C₂alkyl),        aryl(C₀-C₂alkyl), or heteroaryl(C₀-C₂alkyl).

Where each of (ii) and (iii) is substituted with 0 to 3 substituentsindependently chosen from halogen, hydroxy, amino, cyano, nitro, oxo,—COOH, —CONH₂, C₁-C₄alkyl, C₂-C₄alkenyl, C₂-C₄alkynyl, C₁-C₄alkoxy,C₁-C₄alkoxycarbonyl, C₃-C₇cycloalkyl(C₀-C₄carbohydryl),C₃-C₇cycloalkyl(C₀-C₄alkoxy), mono- and di-(C₁-C₄)alkylamino,C₁-C₂haloalkyl, C₁-C₂haloalkoxy, C₂-C₄alkanoyl and phenyl.

A₈ is nitrogen or CR₈.

Wherein, R₈ is hydrogen, halogen, hydroxy, amino, cyano, nitro, or—NHNH₂, or

R₈ is C₁-C₄alkyl, C₁-C₄alkoxy, mono- or di-(C₁-C₄)alkylamino, mono-,di-, or tri-C₁-C₄ alkylhydrazinyl, C₂-C₄alkanoyl, C₁-C₄alkylester,C₁-C₂haloalkyl, or C₁-C₂haloalkoxy; each of which is substituted with 0to 3 substituents independently chosen from hydroxy, amino, halogen,oxo, C₁-C₄alkoxy, C₁-C₂haloalkyl, C₁-C₂haloalkoxy, and mono- anddi-C₁-C₄alkylamino.

R₉ is C₁-C₈alkyl, C₃-C₇cycloalkyl(C₀-C₄alkyl), or phenyl, each of whichis substituted with 0 to 3 substituents independently chosen fromhalogen, hydroxy, amino, cyano, nitro, —COOH, —CONH₂, C₁-C₄alkyl,C₂-C₄alkenyl, C₂-C₄alkynyl, C₁-C₄alkoxy, C₃-C₇cycloalkyl(C₀-C₄alkyl),C₃-C₇cycloalkyl(C₀-C₄alkoxy), mono- and di-(C₁-C₄)alkylaminoC₁-C₂haloalkyl, C₁-C₂haloalkoxy, and C₂-C₄alkanoyl.

The invention includes novel intermediates useful for the synthesis ofantimicrobial compounds of Formula I and Formula II. These intermediatesare compounds of Formula I and Formula II in which R₇ is bromo, iodo,—O(SO₂)CF₃, or —N₂BF₄. The invention provides methods of synthesizingcompounds of Formula I and Formula II comprising coupling anintermediate of the invention to an appropriate aryl or heteroarylboronic acid, aryl or heteroaryl boronic acid ester, or compoundssubstituted with Li, Mg, B, Al, Si, Zn, Cu, Zr, or Sn at the point ofcoupling.

DETAILED DESCRIPTION OF THE INVENTION Chemical Description andTerminology

Prior to setting forth the invention in detail, it may be helpful toprovide definitions of certain terms to be used herein. Compounds of thepresent invention are generally described using standard nomenclature.

In certain situations, the compounds of Formula I and Formula II maycontain one or more asymmetric elements such as stereogenic centers,stereogenic axes and the like, e.g. asymmetric carbon atoms, so that thecompounds can exist in different stereoisomeric forms. These compoundscan be, for example, racemates or optically active forms. For compoundswith two or more asymmetric elements, these compounds can additionallybe mixtures of diastereomers. For compounds having asymmetric centers,it should be understood that all of the optical isomers and mixturesthereof are encompassed. In addition, compounds with carbon-carbondouble bonds may occur in Z- and E-forms, with all isomeric forms of thecompounds being included in the present invention. In these situations,the single enantiomers, i.e., optically active forms can be obtained byasymmetric synthesis, synthesis from optically pure precursors, or byresolution of the racemates. Resolution of the racemates can also beaccomplished, for example, by conventional methods such ascrystallization in the presence of a resolving agent, or chromatography,using, for example a chiral HPLC column.

Where a compound exists in various tautomeric forms, the invention isnot limited to any one of the specific tautomers, but rather includesall tautomeric forms.

The present invention is intended to include all isotopes of atomsoccurring in the present compounds. Isotopes include those atoms havingthe same atomic number but different mass numbers. By way of generalexample, and without limitation, isotopes of hydrogen include tritiumand deuterium and isotopes of carbon include ¹¹C, ¹³C, and ¹⁴C.

Certain compounds are described herein using a general formula thatincludes variables, e.g. A₁, R₂, R₃, R₅, R₆, R₇, A₈, and R₉. Unlessotherwise specified, each variable within such a formula is definedindependently of other variables. Thus, if a group is said to besubstituted, e.g. with 0-2 R*, then said group may be substituted withup to two R* groups and R* at each occurrence is selected independentlyfrom the definition of R*. Also, combinations of substituents and/orvariables are permissible only if such combinations result in stablecompounds.

The term “substituted”, as used herein, means that any one or morehydrogens on the designated atom or group is replaced with a selectionfrom the indicated group, provided that the designated atom's normalvalence is not exceeded. When a substituent is oxo (i.e., ═O), imine(e.g. ═NHR), or oxime (e.g. ═NOR) then 2 hydrogens on the atom arereplaced. An “oxo,” imine, or oxime substituent on an aromatic group orheteroaromatic group destroys the aromatic character of that group, e.g.a pyridyl substituted with oxo is pyridone. Combinations of substituentsand/or variables are permissible only if such combinations result instable compounds or useful synthetic intermediates. A stable compound orstable structure is meant to imply a compound that is sufficientlyrobust to survive isolation from a reaction mixture, and subsequentformulation into an effective therapeutic agent. Unless otherwisespecified substituents are named into the core structure. For example,it is to be understood that when cycloalkyl(alkyl) is listed as apossible substituent the point of attachment of this substituent to thecore structure is in the alkyl portion.

A dash (“—”) that is not between two letters or symbols is used toindicate a point of attachment for a substituent. For example, —CONH₂ isattached through the carbon atom.

As used herein, “alkyl” is intended to include both branched andstraight-chain saturated aliphatic hydrocarbon groups, having thespecified number of carbon atoms. Thus, the term C₁-C₆ alkyl as usedherein includes alkyl groups having from 1 to about 6 carbon atoms. WhenC₀-C_(n) alkyl is used herein in conjunction with another group, forexample, arylC₀-C₄alkyl, the indicated group, in this case aryl, iseither directly bound by a single covalent bond (C₀), or attached by analkyl chain having the specified number of carbon atoms, in this casefrom 1 to about 4 carbon atoms. Examples of alkyl include, but are notlimited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl,n-pentyl, and sec-pentyl. Preferred alkyl groups are lower alkyl groups,those alkyl groups having from 1 to about 8 carbon atoms, e.g. C₁-C₈ andC₁-C₆alkyl groups.

“Alkenyl” as used herein, indicates a hydrocarbon chain of either astraight or branched configuration having one or more carbon-carbondouble bond bonds, which may occur at any stable point along the chain.Examples of alkenyl groups include ethenyl and propenyl.

“Alkynyl” as used herein, indicates a hydrocarbon chain of either astraight or branched configuration having one or more triplecarbon-carbon bonds that may occur in any stable point along the chain,such as ethynyl and propynyl.

“Carbohydryl” as used herein, includes both branched and straight-chainhydrocarbon groups, which are saturated or unsaturated, having thespecified number of carbon atoms. When C₀-C_(n) carbohydrylis usedherein in conjunction with another group, for example,arylC₀-C₄carbohydryl, the indicated group, in this case aryl, is eitherdirectly bound by a single covalent bond (C₀), or attached by ancarbohydryl chain, such as an alkyl chain, having the specified numberof carbon atoms, in this case from 1 to about 4 carbon atoms. Examplesinclude C₁-C₆alkyl, such as methyl, or 5-butyl, C₂-C₆alkynyl such andhexynyl, and C₂-C₆ alkenyl, such as 1-propenyl.

“Alkoxy” represents an alkyl group as defined above with the indicatednumber of carbon atoms attached through an oxygen bridge. Examples ofalkoxy include, but are not limited to, methoxy, ethoxy, n-propoxy,propoxy, n-butoxy,2-butoxy, t-butoxy, n-pentoxy, 2-pentoxy, 3-pentoxy,isopentoxy, neopentoxy, n-hexoxy, 2-hexoxy, 3-hexoxy, and3-methylpentoxy.

“Alkanoyl” indicates an alkyl group as defined above, attached through aketo (—(C═O)—) bridge. Alkanoyl groups have the indicated number ofcarbon atoms, with the carbon of the keto group being included in thenumbered carbon atoms. For example a C2alkanoyl group is an acetyl grouphaving the formula CH₃(C═O)—.

As used herein, the terms “mono- or di-alkylamino” or “mono- anddi-alkylamino” indicate secondary or tertiary alkyl amino groups,wherein the alkyl groups are as defined above and have the indicatednumber of carbon atoms. The point of attachment of the alkylamino groupis on the nitrogen. Examples of mono- and di-alkylamino groups includeethylamino, dimethylamino, and methyl-propyl-amino.

The term “mono- or di-alkylcarbamate” indicates 1 or 2 independentlychosen alkyl groups, as define above, attached through a carbamate(—O(C═O)NRR) linkage where R represents the alkyl groups.Mono-alkylcarbamate groups have the formula (—O(C═O)NHR.

The term “alkylester” indicates and alkyl group as define above attachedthrough an ester linkage, i.e. a group of the formula —O(C═O)alkyl.

The term “mono-, di-, or tri-alkylhydrazinyl” indicates from 1 to 3independently chosen alkyl group as defined above attached through asingle-bonded nitrogen-nitrogen linkage. At least one of the alkylgroups is attached to the terminal nitrogen (the nitrogen not bound tothe core structure). When the term mono- or di-alkylhydrazinyl is usedonly the terminal nitrogen is alkyl substituted. Examples ofalkylhydrazinyl groups include 2-butyl-1-hydrazinyl,2-butyl-2-methyl-1-hydrazinyl, and 1,2-dimethyl-2-propyl-1-hydrazinyl.

The term “alkylthio” indicates an alkyl group as defined above attachedthrough a sulfur linkage, i.e. a group of the formula alkyl-S—. Examplesinclude ethylthio and pentylthio.

As used herein, the term “aryl” indicates aromatic groups containingonly carbon in the aromatic ring or rings. Typical aryl groups contain 1to 3 separate, fused, or pendant rings and from 6 to about 18 ringatoms, without heteroatoms as ring members. When indicated, such arylgroups may be further substituted with carbon or non-carbon atoms orgroups. Such substitution may include fusion to a 5 to 7-memberedsaturated cyclic group that optionally contains 1 or 2 heteroatomsindependently chosen from N, O, and S, to form, for example, a3,4-methylenedioxy-phenyl group. Aryl groups include, for example,phenyl, naphthyl, including 1-naphthyl and 2-naphthyl, and bi-phenyl.

In the term “aryl(alkyl)”, aryl and alkyl are as defined above, and thepoint of attachment is on the alkyl group. This term encompasses, but isnot limited to, benzyl, phenylethyl, and piperonyl. Likewise, in theterm aryl(carbohydryl), aryl and carbohydryl are as defined above andthe point of attachment is on the carbohydryl group, for example aphenylpropen-1-yl group.

“Cycloalkyl” as used herein, indicates saturated hydrocarbon ringgroups, having the specified number of carbon atoms, usually from 3 toabout 8 ring carbon atoms, or from 3 to about 7 carbon atoms. Examplesof cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, orcyclohexyl as well as bridged or caged saturated ring groups such asnorborane or adamantane.

“Cycloalkenyl” as used herein, indicates an unsaturated, but notaromatic, hydrocarbon ring having at least one carbon-carbon doublebond. Cycloalkenyl groups contain from 4 to about 8 carbon atoms,usually from 4 to about 7 carbon atoms. Examples include cyclohexenyland cyclobutenyl.

In the terms “cycloalkyl(alkyl),” “cycloalkyl(carbohydryl),” and“cycloalkyl(alkoxy)” the terms cycloalkyl, alkyl, carbohydryl, andalkoxy are as defined above, and the point of attachment is on thealkyl, carbohydryl, or alkoxy group respectively. These terms includeexamples such as cyclopropylmethyl, cyclohexylmethyl,cyclohexylpropenyl, and cyclopentylethyoxy.

In the terms “cycloalkenyl(alkyl)” “cycloalkenyl(carbohydryl)” and theterms cycloalkenyl, alkyl, and carbohydryl are as defined above, and thepoint of attachment is on the alkyl or carbohydryl group respectively.These terms include examples such as cyclobutenylmethyl,cyclohexenylmethyl, and cyclohexylpropenyl.

“Haloalkyl” indicates both branched and straight-chain saturatedaliphatic hydrocarbon groups having the specified number of carbonatoms, substituted with 1 or more halogen atoms, generally up to themaximum allowable number of halogen atoms. Examples of haloalkylinclude, but are not limited to, trifluoromethyl, difluoromethyl,2-fluoroethyl, and penta-fluoroethyl.

“Haloalkoxy” indicates a haloalkyl group as defined above attachedthrough an oxygen bridge.

“Halo” or “halogen” as used herein refers to fluoro, chloro, bromo, oriodo.

As used herein, “heteroaryl” indicates a stable 5- to 7-memberedmonocyclic or 7- to 10-membered bicyclic heterocyclic ring whichcontains at least 1 aromatic ring that contains from 1 to 4, orpreferably from 1 to 3, heteroatoms chosen from N, O, and S, withremaining ring atoms being carbon. When the total number of S and Oatoms in the heteroaryl group exceeds 1, these heteroatoms are notadjacent to one another. It is preferred that the total number of S andO atoms in the heteroaryl group is not more than 2. It is particularlypreferred that the total number of S and O atoms in the aromaticheterocycle is not more than 1. A nitrogen atom in a heteroaryl groupmay optionally be quaternized. When indicated, such heteroaryl groupsmay be further substituted with carbon or non-carbon atoms or groups.Such substitution may include fusion to a 5 to 7-membered saturatedcyclic group that optionally contains 1 or 2 heteroatoms independentlychosen from N, O, and S, to form, for example, a[1,3]dioxolo[4,5-c]pyridyl group. Examples of heteroaryl groups include,but are not limited to, pyridyl, indolyl, pyrimidinyl, pyridizinyl,pyrazinyl, imidazolyl, oxazolyl, furanyl, thiophenyl, thiazolyl,triazolyl, tetrazolyl, isoxazolyl, quinolinyl, pyrrolyl, pyrazolyl,benz[b]thiophenyl, isoquinolinyl, quinazolinyl, quinoxalinyl, thienyl,isoindolyl, and 5,6,7,8-tetrahydroisoquinoline.

In the terms “heteroarylalkyl” and “heteroaryl(carbohydryl),”heteroaryl, alkyl, and carbohydryl are as defined above, and the pointof attachment is on the alkyl or carbohydryl group respectively. Theseterms include such examples as pyridylmethyl, thiophenylmethyl, andpyrrolyl(1-ethyl).

The term “heterocycloalkyl” indicates a saturated cyclic groupcontaining from 1 to about 3 heteroatoms chosen from N, O, and S, withremaining ring atoms being carbon. Heterocycloalkyl groups have from 3to about 8 ring atoms, and more typically have from 5 to 7 ring atoms. AC₂-C₇heterocycloalkyl group contains from 2 to about 7 carbon ring atomsand at least one ring atom chosen from N, O, and S. Examples ofheterocycloalkyl groups include morpholinyl, piperazinyl, piperidinyl,and pyrrolidinyl groups. A nitrogen in a heterocycloalkyl group mayoptionally be quaternized.

The term “heterocyclic group” indicates a 5-6 membered saturated,partially unsaturated, or aromatic ring containing from 1 to about 4heteroatoms chosen from N, O, and S, with remaining ring atoms beingcarbon or a 7-10 membered bicyclic saturated, partially unsaturated, oraromatic heterocylic ring system containing at least 1 heteroatom in thetwo ring system chosen from N, O, and S and containing up to about 4heteroatoms independently chosen from N, O, and S in each ring of thetwo ring system. Unless otherwise indicated, the heterocyclic ring maybe attached to its pendant group at any heteroatom or carbon atom thatresults in a stable structure. When indicated the heterocyclic ringsdescribed herein may be substituted on carbon or on a nitrogen atom ifthe resulting compound is stable. A nitrogen atom in the heterocycle mayoptionally be quaternized. It is preferred that the total number ofheteroatoms in a heterocyclic groups is not more than 4 and that thetotal number of S and O atoms in a heterocyclic group is not more than2, more preferably not more than 1. Examples of heterocyclic groupsinclude, pyridyl, indolyl, pyrimidinyl, pyridizinyl, pyrazinyl,imidazolyl, oxazolyl, furanyl, thiophenyl, thiazolyl, triazolyl,tetrazolyl, isoxazolyl, quinolinyl, pyrrolyl, pyrazolyl,benz[b]thiophenyl, isoquinolinyl, quinazolinyl, quinoxalinyl, thienyl,isoindolyl, dihydroisoindolyl, 5,6,7,8-tetrahydroisoquinoline,pyridinyl, pyrimidinyl, furanyl, thienyl, pyrrolyl, pyrazolyl,pyrrolidinyl, morpholinyl, piperazinyl, piperidinyl, and pyrrolidinyl.

Additional examples heterocyclic groups include, but are not limited to,phthalazinyl, oxazolyl, indolizinyl, indazolyl, benzothiazolyl,benzimidazolyl, benzofuranyl, benzoisoxolyl, dihydro-benzodioxinyl,oxadiazolyl, thiadiazolyl, triazolyl, tetrazolyl, oxazolopyridinyl,imidazopyridinyl, isothiazolyl, naphthyridinyl, cinnolinyl, carbazolyl,beta-carbolinyl, isochromanyl, chromanonyl, chromanyl,tetrahydroisoquinolinyl, isoindolinyl, isobenzotetrahydrofuranyl,isobenzotetrahydrothienyl, isobenzothienyl, benzoxazolyl,pyridopyridinyl, benzotetrahydrofuranyl, benzotetrahydrothienyl,purinyl, benzodioxolyl, triazinyl, phenoxazinyl, phenothiazinyl,pteridinyl, benzothiazolyl, imidazopyridinyl, imidazothiazolyl,dihydrobenzisoxazinyl, benzisoxazinyl, benzoxazinyl,dihydrobenzisothiazinyl, benzopyranyl, benzothiopyranyl, coumarinyl,isocoumarinyl, chromanyl, tetrahydroquinolinyl, dihydroquinolinyl,dihydroquinolinonyl, dihydroisoquinolinonyl, dihydrocoumarinyl,dihydroisocoumarinyl, isoindolinonyl, benzodioxanyl, benzoxazolinonyl,pyrrolyl N-oxide, pyrimidinyl N-oxide, pyridazinyl N-oxide, pyrazinylN-oxide, quinolinyl N-oxide, indolyl N-oxide, indolinyl N oxide,isoquinolyl N-oxide, quinazolinyl N-oxide, quinoxalinyl N-oxide,phthalazinyl N-oxide, imidazolyl N-oxide, isoxazolyl N-oxide, oxazolylN-oxide, thiazolyl N-oxide, indolizinyl N oxide, indazolyl N-oxide,benzothiazolyl N-oxide, benzimidazolyl N-oxide, pyrrolyl N-oxide,oxadiazolyl N-oxide, thiadiazolyl N-oxide, tetrazolyl N-oxide,benzothiopyranyl S-oxide, and benzothiopyranyl S,S-dioxide.

As used herein “Active agents” are compounds that have pharmaceuticalutility, e.g. may be used to treat a patient suffering from a disease orcondition, or may be used prophylacticly to prevent the onset of adisease or condition in a patient, or that may be used to enhance thepharmaceutical activity of other compounds.

“Salts” of the compounds of the present invention include inorganic andorganic acid and base addition salts. The salts of the present compoundscan be synthesized from a parent compound that contains a basic oracidic moiety by conventional chemical methods. Generally, such saltscan be prepared by reacting free acid forms of these compounds with astoichiometric amount of the appropriate base (such as Na, Ca, Mg, or Khydroxide, carbonate, bicarbonate, or the like), or by reacting freebase forms of these compounds with a stoichiometric amount of theappropriate acid. Such reactions are typically carried out in water orin an organic solvent, or in a mixture of the two. Generally,non-aqueous media like ether, ethyl acetate, ethanol, isopropanol, oracetonitrile are preferred, where practicable. Salts of the presentcompounds further include solvates of the compounds and of the compoundsalts.

“Pharmaceutically acceptable salts” includes derivatives of thedisclosed compounds wherein the parent compound is modified by makingnon-toxic acid or base salts thereof, and further refers topharmaceutically acceptable solvates of such compounds and such salts.Examples of pharmaceutically acceptable salts include, but are notlimited to, mineral or organic acid salts of basic residues such asamines; alkali or organic salts of acidic residues such as carboxylicacids; and the like. The pharmaceutically acceptable salts include theconventional non-toxic salts and the quaternary ammonium salts of theparent compound formed, for example, from non-toxic inorganic or organicacids. For example, conventional non-toxic acid salts include thosederived from inorganic acids such as hydrochloric, hydrobromic,sulfuric, sulfamic, phosphoric, nitric and the like; and the saltsprepared from organic acids such as acetic, propionic, succinic,glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic,maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic,mesylic, esylic, besylic, sulfanilic, 2-acetoxybenzoic, fumaric,toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic,HOOC—(CH₂)_(n)—COOH where n is 0-4, and the like. Lists of additionalsuitable salts may be found, e.g., in Remington's PharmaceuticalSciences, 17th ed., Mack Publishing Company, Easton, Pa., p. 1418(1985).

The term “prodrugs” includes any compounds that become compounds ofFormula I when administered to a mammalian subject, e.g., upon metabolicprocessing of the prodrug. Examples of prodrugs include, but are notlimited to, acetate, formate and benzoate and like derivatives offunctional groups (such as alcohol or amine groups) in the compounds ofFormula I and Formula II.

The term “therapeutically effective amount” of a compound of thisinvention means an amount effective, when administered to a human ornon-human patient, to provide a therapeutic benefit such as anamelioration of symptoms, e.g., an amount effective to decrease thesymptoms of a microbial infection, and preferably an amount sufficientto reduce the symptoms of a bacterial, fungal, or protozoal infection.In certain circumstances a patient suffering from a microbial infectionmay not present symptoms of being infected. Thus a therapeuticallyeffective amount of a compound is also an amount sufficient to prevent asignificant increase or significantly reduce the detectable level ofmicroorganism or antibodies against the microorganism in the patient'sblood, serum, other bodily fluids, or tissues. The invention alsoincludes using compounds of Formula I and Formula II in prophylactictherapies. In the context of prophylactic or preventative treatment a“therapeutically effective amount” is an amount sufficient tosignificantly decrease the treated animal's risk of contracting amicroorganism infection. A significant reduction is any detectablenegative change that is statistically significant in a standardparametric test of statistical significance such as Student's T-test,where p<0.05.

Antimicrobial Compounds

For the purposes of this document, the following numbering system willapply to the core 9H-isothiazolo[5,4-b]quinoline-3,4-dione (whenA₁=sulfur) structure or core 9H-isoxazolo[5,4-b]quinoline-3,4-dione(when A₁=oxygen) structure. The numbers 1 through 9 refer specificallyto positions within the tricyclic ring system whereas the letters A, Band C refer to the specific six (rings A and B) or five (ring C) memberrings as shown below.

In addition to the compounds of Formula I and Formula II, describedabove the invention also includes compounds of Formula I and Formula IIin which the variables (e.g. A₁, R₂, R₃, R₄, etc.) carry definitionsother than those set forth above.

The A₁ Variable

In certain embodiments, the invention includes compounds of Formula Iand Formula II A₁ is Sulfur.

In other embodiments A₁ is SO.

The invention also includes compounds of Formula I and Formula II inwhich A₁ is SO₂.

In still other embodiments A₁ is O.

The R₂ Variable

The invention includes compounds of Formula I in which R₂ is hydrogen orR₂ is C₁-C₆alkyl or C₃-C₇cycloalkyl(C₀-C₄alkyl), each of which issubstituted with at least one substituent chosen from hydroxy, amino,—COOH, —(C═O)NR₁₀OR₁₁, and —CONH₂; and is substituted with 0 to 3substituents independently chosen from halogen, hydroxy, amino, cyano,nitro, —COOH, —CONH₂, C₁-C₄alkyl, C₁-C₄alkoxy, C₁-C₂haloalkyl,C₁-C₂haloalkoxy, and mono- and di-C₁-C₄alkylamino, and C₂-C₄alkanoyl.Certain embodiments of the invention pertain to compounds of Formula Iin which R₂ is hydrogen.

The R₃ Variable

The invention includes compounds and salts of Formula II in which R₃ isC₁-C₆alkyl, C₁-C₆alkanoyl, mono- or di-C₁-C₆alkylcarbamate, orC₁-C₆alkylsulfonate; each of which is substituted with 0 to 3substituents independently chosen from halogen, hydroxy, amino, cyano,nitro, C₁-C₄alkoxy, mono- and di-C₁-C₄alkylamino, C₁-C₂haloalkyl, andC₁-C₂haloalkoxy.

The invention also includes compounds and salts of Formula II in whichR₃ is C₁-C₆alkyl or C₁-C₆alkanoyl, each of which is substituted with 0to 3 substituents independently chosen from halogen, hydroxy, amino,cyano, C₁-C₂alkoxy, mono- and di-C₁-C₂alkylamino, C₁-C₂haloalkyl, andC₁-C₂haloalkoxy.

The invention also includes compounds and salts of Formula II in whichR3 is C₁-C₆alkyl or C₁-C₆alkanoyl.

The R₅ Variable

Certain embodiments of the invention pertain to compounds and salts ofFormula I and Formula II in which R₅ is hydrogen, amino, C₁-C₂alkyl,C₁-C₂alkoxy, mono- or di-(C₁-C₄)alkylamino, or mono- or di-C₁-C₄alkylhydrazinyl.

The invention also includes compounds and salts of Formula I and FormulaII in which R₅ hydrogen, amino, mono- or di-(C₁-C₂)alkylamino, or mono-or di-C₁-C₂ alkylhydrazinyl.

The invention includes compounds and salts of Formula I and Formula IIin which R₅ is hydrogen.

The R₆ Variable

The invention includes compounds and salts of Formula I and Formula IIin which R₆ is hydrogen, halogen, or amino.

In certain embodiments the invention pertains to compounds and salts ofFormula I and Formula II in which R₆ is fluoro or hydrogen.

The invention includes compounds and salts of Formula I and Formula IIin which R₆ is halogen and R₈ is C₁-C₂alkoxy.

The invention includes compounds and salts of Formula I and Formula IIin which R₆ is fluorine and R₈ is methoxy.

The R₇ Variable

The invention includes compounds and salts of Formula I and Formula IIin which R₇ is bromo, iodo, —O(SO₂)CF₃, or —N₂BF₄. These compounds areparticularly useful as intermediates in the synthesis of antimicrobialcompounds of Formula I and Formula II.

The invention includes compounds and salts of Formula I and Formula IIin which R₇ is XR_(A) where X is absent, CH₂—CH₂—, —CH═CH—, or —C═C—;and R_(A) is a 7-10 membered bicyclic saturated, partially unsaturated,or aromatic carbocyclic group, or R_(A) is a 5-6 membered saturated,partially unsaturated, or aromatic heterocylic group bound via a carbonatom when X is absent or —CH₂—CH₂—, or bound via a carbon or nitrogenatom when X is —CH═CH— or —C≡C— or a R_(A) is a 7-10 membered bicyclicsaturated, partially unsaturated, or aromatic heterocylic group boundvia a carbon atom when X is absent or —CH₂—CH₂—, or bound via a carbonor nitrogen atom when X is —CH═CH— or —C═C—; each of which R_(A) issubstituted with 0 to 5 substituents independently chosen from (i),(ii), and (iii). where:

-   -   (i) is chosen from halogen, hydroxy, amino, cyano, and nitro;        and    -   (ii) is chosen from C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl,        mono- and di-(C₁-C₄)alkylamino, C₁-C₂haloalkyl, C₁-C₂haloalkoxy,        C₃-C₇cycloalkyl(C₀-C₄alkyl), C₃-C₇cycloalkyl(C₀-C₄alkoxy),        C₄-C₇cycloalkenyl(C₀-C₄alkyl), aryl(C₀-C₆carbohydryl),        aryl(C₁-C₄alkoxy), C₂-C₆heterocycloalkyl(C₀-C₄alkyl),        heteroaryl(C₀-C₆carbohydryl), C₁-C₆alkylthio, ═NOR₁₀,        —(C₀-C₄alkyl)(C═O)R₁₀, —(C₀-C₄alkyl)O(C═O)R₁₀,        —(C₀-C₄alkyl)(C═O)NR₁₀R₁₁, —(C₀-C₄alkyl)O(C═O)NR₁₀R₁₁,        —(C₀-C₄alkyl) (C═O)OR₁₀, —(C₀-C₄alkyl)NR₁₀(C═O)R₁₁,        —(C₀-C₄alkyl)NR₁₀(C═O)OR₁₁, —(C₀-C₄alkyl)NR₁₀(C═O)NR₁₁R₁₂,        —(C_(O)-C₄alkyl)NR₁₀(C═O)(C₁-C₄alkyl)NR₁₁(C═O)O—R₁₂,        —(C₀-C₄alkyl)NR₁₀(C═S)NR₁₁R₁₂, —(C₀-C₄alkyl)NR₁₀NR₁₁R₁₂,        —(C₀-C₄alkyl)N═NR₁₃, —(C₀-C₄alkyl)SO₃R₁₀,        —(C₀-C₄alkyl)(S═O)OR₁₀, —(C₀-C₄alkyl)SO₂R₁₃,        —(C₀-C₄alkyl)SO₂NR₁₀R₁₁, and —(C₀-C₄alkyl)NR₁₀SO₂R₁₃; and    -   (iii) is chosen from —OR_(D), —(C═O)R_(D), —SO₂R_(D), —SO₃R_(D),        —NR₁₀SO₂R_(D), where R_(D) is C₁-C₄alkyl,        C₃-C₇cycloalkyl(C₀-C₂alkyl), C₂-C₆heterocycloalkyl(C₀-C₂alkyl),        aryl(C₀-C₂alkyl), or heteroaryl(C₀-C₂alkyl).

Where each of (ii) and (iii) is substituted with 0 to 3 substituentsindependently chosen from halogen, hydroxy, amino, cyano, nitro, oxo,—COOH, —CONH₂, C₂-C₄alkenyl, C₂-C₄alkynyl, C₁-C₄alkoxy,C₁-C₄alkoxycarbonyl, C₃-C₇cycloalkyl(C₀-C₄alkyl),C₃-C₇cycloalkyl(C₀-C₄alkoxy), mono- and di-(C₁-C₄)alkylamino,C₁-C₂haloalkyl, C₁-C₂haloalkoxy, C₂-C₄alkanoyl, and phenyl.

The invention also pertains to compounds and salts of Formula I andFormula II in which

R₇ is XR_(A), X is absent, CH₂—CH₂—, —CH═CH—, or —C≡—; and

R_(A) is naphthyl, dihydronapthyl, tetrahydronapthyl, pyridyl,pyrimidinyl, pyrazinyl, furanyl, benz[b]thiophenyl, benzofuranyl,quinolinyl, isoquinolinyl, quinazolinyl, quinoxalinyl, isoxazolyl,indolyl, dihydroindolyl, pyrrolyl, pyrazolyl, imidazolyl, thienyl,isoindolyl, dihydroisoindolyl, tetrahydropyridinyl,tetrahydroisoquinolinyl, or piperidin-4-yl group; each of which issubstituted with 0 to 5 substituents independently chosen from (i),(ii), and (iii) which are as defined above.

The invention also includes compounds and salts of Formula I and FormulaII in which

R₇ is XR_(A), X is absent, CH₂—CH₂—, —CH═CH—, or —C≡C—; and

R_(A) is naphthyl, dihydronapthyl, tetrahydronapthyl, pyridyl,pyrimidinyl, pyrazinyl, furanyl, benz[b]thiophenyl, benzofuranyl,quinolinyl, isoquinolinyl, quinazolinyl, quinoxalinyl, isoxazolyl,indolyl, dihydroindolyl, pyrrolyl, pyrazolyl, imidazolyl, thienyl,isoindolyl, dihydroisoindolyl, tetrahydropyridinyl,tetrahydroisoquinolinyl, or piperidin-4-yl group; each of which issubstituted with 0 to 5 substituents independently chosen from (i),(ii), and (iii). In this embodiment (i), (ii), and (iii) carry thefollowing definitions:

-   -   (i) is chosen from halogen, hydroxy, amino, cyano, and nitro,    -   (ii) is chosen from C₁-C₄alkyl, C₂-C₄alkenyl, C₁-C₄alkoxy, mono-        and di-(C₁-C₄)alkylamino, C₁-C₂haloalkyl, C₁-C₂haloalkoxy,        C₃-C₇cycloalkyl(C₀-C₂alkyl), C₃-C₇cycloalkyl(C₀-C₂alkoxy),        phenyl(C₀-C₂alkyl), phenyl(C₀-C₂alkoxy),        pyrrolidinyl(C₀-C₂alkyl), piperidinyl(C₀-C₂alkyl),        piperazinyl(C₀-C₂alkyl), morpholinyl(C₀-C₂alkyl),        thiomorpholinyl(C₀-C₂alkyl), pyridyl, pyrimidinyl, pyrazinyl,        furanyl, benzofuranyl, pyrrolyl, pyrazolyl, imidazolyl, thienyl,        C₁-C₄alkylthio, ═NOR₁₀, —(C₀-C₄alkyl)(C═O)R₁₀,        —(C₀-C₄alkyl)O(C═O)R₁₀, —(C₀-C₄alkyl)(C═O)NR₁₀R₁₁,        —(C₀-C₄alkyl)O(C═O)NR₁₀R₁₁, —(C₀-C₄alkyl) (C═O)OR₁₀,        —(C₀-C₄alkyl)NR₁₀(C═O)R₁₁, —(C₀-C₄alkyl)NR₁₀(C═O)OR₁₁,        —(C₀-C₄alkyl)NR₁₀(C═O)NR₁₁R₁₂,        —(C₀-C₄alkyl)NR₁₀(C═O)(C₁-C₄alkyl)NR₁₁(C═O)O—R₁₂,        —(C₀-C₄alkyl)NR₁₀(C═S)NR₁₁R₁₂, —(C₀-C₄alkyl)NR₁₀NR₁₁R₁₂,        —(C₀-C₄alkyl)N═NR₁₃, —(C₀-C₄alkyl)SO₃R₁₀,        —(C₀-C₄alkyl)(S═O)OR₁₀, —(C₀-C₄alkyl)SO₂R₁₃,        —(C₀-C₄alkyl)SO₂NR₁₀R₁₁, and —(C₀-C₄alkyl)NR₁₀SO₂R₁₃; and    -   (iii) is chosen from —OR_(D), —(C═O)R_(D), —SO₂R_(D), —SO₃R_(D),        —NR₁₀SO₂R_(D), where R_(D) is C₁-C₄alkyl,        C₃-C₇cycloalkyl(C₀-C₂alkyl), pyrrolidinyl(C₀-C₂alkyl),        piperidinyl(C₀-C₂alkyl), piperazinyl(C₀-C₂alkyl),        morpholinyl(C₀-C₂alkyl), thiomorpholinyl(C₀-C₂alkyl),        phenyl(C₀-C₂alkyl), naphthyl(C₀-C₂alkyl), pyridyl(C₀-C₂alkyl),        pyrimidinyl(C₀-C₂alkyl), pyrazinyl(C₀-C₂alkyl),        furanyl(C₀-C₂alkyl), benz[b]thiophenyl(C₀-C₂alkyl),        benzofuranyl(C₀-C₂alkyl), quinolinyl(C₀-C₂alkyl),        isoquinolinyl(C₀-C₂alkyl), quinazolinyl(C₀-C₂alkyl),        isoxazolyl(C₀-C₂alkyl), indolyl(C₀-C₂alkyl),        dihydroindolyl(C₀-C₂alkyl), pyrrolyl(C₀-C₂alkyl),        pyrazolyl(C₀-C₂alkyl), imidazolyl(C₀-C₂alkyl),        thienyl(C₀-C₂alkyl), isoindolyl(C₀-C₂alkyl), or        dihydroisoindolyl(C₀-C₂alkyl).

Where each of (ii) and (iii) is substituted with 0 to 3 substitutentsindependently chosen from halogen, hydroxy, amino, cyano, nitro, oxo,—COOH, —CONH₂, C₁-C₄alkyl, C₂-C₄alkenyl, C₂-C₄alkynyl, C₁-C₄alkoxy,C₁-C₄alkoxycarbonyl, C₃-C₇cycloalkyl(C₀-C₄alkyl),C₃-C₇cycloalkyl(C₀-C₄alkoxy), mono- and di-(C₁-C₄)alkylamino,C₁-C₂haloalkyl, C₁-C₂haloalkoxy, C₂-C₄alkanoyl, and phenyl.

The invention further includes compounds and salts of Formula I andFormula II in which R₇ is XR_(A), X is absent, CH₂—CH₂—, —CH═CH—, or—C≡C—; and R_(A) is a naphthyl, dihydronapthyl, tetrahydronapthyl,pyridyl, pyrimidinyl, pyrazinyl, furanyl, benz[b]thiophenyl,benzofuranyl, quinolinyl, isoquinolinyl, quinazolinyl, quinoxalinyl,isoxazolyl, indolyl, dihydroindolyl, pyrrolyl, pyrazolyl, imidazolyl,thienyl, isoindolyl, dihydroisoindolyl, tetrahydropyridinyl,tetrahydroisoquinolinyl, or piperidin-4-yl group; each of which issubstituted with 0 to 5 substituents independently chosen from (i),(ii), and (iii).

In this embodiment (i) is chosen from halogen, hydroxy, amino, cyano,and nitro, (ii) is chosen from C₁-C₄alkyl, C₂-C₄alkenyl, C₁-C₄alkoxy,mono- and di-(C₁-C₄)alkylamino, C₁-C₂haloalkyl, C₁-C₂haloalkoxy,C₃-C₇cycloalkyl(C₀-C₂alkyl), C₃-C₇cycloalkyl(C₀-C₂alkoxy),phenyl(C₀-C₂alkyl), phenyl(C₀-C₂alkoxy), pyrrolidinyl, piperidinyl,piperazinyl, morpholinyl, thiomorpholinyl, pyridyl, thienyl,C₁-C₄alkylthio, —(C═O)OR₁₀, and —(C═O)NR₁₀R₁₁; and (iii) is chosen from—OR_(D), —(C═O)R_(D), —SO₂R_(D), —SO₃R_(D), and —NR₁₀SO₂R_(D), whereR_(D) is C₁-C₄alkyl, C₃-C₇cycloalkyl(C₀-C₂alkyl), piperidinyl,piperazinyl, phenyl, naphthyl, or pyridyl.

Each of (ii) and (iii) is substituted with 0 to 3 substituentsindependently chosen from halogen, hydroxy, amino, cyano, nitro, oxo,—COOH, —CONH₂, C₁-C₄alkyl, C₁-C₄alkoxy, C₁-C4alkoxycarbonyl,C₃-C₇cycloalkyl(C₀-C₂alkyl), mono- and di-(C₁-C₄)alkylamino,C₁-C₂haloalkyl, C₁-C₂haloalkoxy, C₂-C₄alkanoyl, and phenyl.

Another embodiment of the invention pertains to compounds and salts ofFormula I and Formula II in which R₇ is XR_(A), X is absent, CH₂—CH₂—,—CH═CH—, or —C═C—; and R_(A) is a naphthyl, dihydronapthyl,tetrahydronapthyl, pyridyl, pyrimidinyl, pyrazinyl, furanyl,benz[b]thiophenyl, benzofuranyl, quinolinyl, isoquinolinyl,quinazolinyl, quinoxalinyl, isoxazolyl, indolyl, dihydroindolyl,pyrrolyl, pyrazolyl, imidazolyl, thienyl, isoindolyl, dihydroisoindolyl,tetrahydropyridinyl tetrahydroisoquinolinyl, or piperidin-4-yl group;each of which is substituted with 0 to 5 substituents independentlychosen from halogen, hydroxy, amino, cyano, nitro, C₁-C₄alkyl,C₂-C₄alkenyl, C₁-C₄alkoxy, mono- and di-(C₁-C₄)alkylamino,C₁-C₂haloalkyl, C₁-C₂haloalkoxy, C₃-C₇cycloalkyl(C₀-C₂alkyl),C₃-C₁cycloalkyl(C₀-C₂alkoxy), phenyl(C₀-C₂alkyl), phenyl(C₀-C₂alkoxy),pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl,pyridyl, thienyl, C₁-C₄alkylthio, —(C═O)OR₁₀, —(C═O)NR₁₀R₁₁; —OR_(D),—(C═O)R_(D), —SO₂R_(D), —SO₃R_(D), and —NR₁₀SO₂R_(D).

In this embodiment R_(D) is C₁-C₄alkyl, piperidinyl, phenyl, naphthyl,or pyridyl; and each R_(D) is substituted with 0 to 3 substituentsindependently chosen from halogen, hydroxy, amino, cyano, C₁-C₂alkyl,C₁-C₂alkoxy, mono- and di-(C₁-C₂)alkylamino, C₁-C₂haloalkyl, andC₁-C₂haloalkoxy.

The invention includes certain compounds of Formula I and Formula II inwhich R₇ is XR_(A) and X is absent.

Other embodiments of the invention pertain to compounds and salts ofFormula I and Formula II in which R₇ is XR_(A), X is absent, CH₂—CH₂—,—CH═CH—, or —C≡C—; and R_(A) is a pyridyl, pyrimidinyl, furanyl,quinolinyl, indolyl, pyrrolyl, isoindolyl, tetrahydroisoquinolinyl, orthienyl group; each of which is substituted with 0 to 3 substituentsindependently chosen from halogen, hydroxy, amino, cyano, C₁-C₄alkyl,C₁-C₄alkoxy, mono- and di-(C₁-C₄)alkylamino, C₁-C₂haloalkyl, andC₁-C₂haloalkoxy.

The invention also includes certain compounds and salt of Formula I andFormula II in which R_(A) is pyrid-3-yl, pyrid-4-yl, pyrimidin-5-yl,furan-3-yl, quinolin-3-yl, quinolin-5-yl, quinolin-6-yl, isoindol-5-yl,tetrahydroisoquinolin-5-yl, tetrahydroisoquinolin-6-yl,tetrahydroisoquinolin-7-yl, tetrahydroisoquinolin-8-yl, or indol-5-yl,each of which is substituted with 0 to 2 substituents independentlychosen from halogen, hydroxy, amino, C₁-C₂alkyl, and C₁-C₂alkoxy.

Further included herein are certain compounds and salts of Formula I andFormula II in which R_(A) is pyridyl-3-yl or pyrid-4-yl, each of whichis substituted with 1 or 2 substituents independently chosen fromfluoro, amino, hydroxy, cyano, and methyl.

The invention also provides compounds and salts of Formula I and FormulaII in which R_(A) is tetrahydroisoquinolin-5-yl,tetrahydroisoquinolin-6-yl, tetrahydroisoquinolin-7-yl, ortetrahydroisoquinolin-8-yl, each of which is substituted with 0 to 3substituents independently chosen from C₁-C₃alkyl.

Also included are compounds and salts of Formula I and Formula II inwhich R_(A) is tetrahydroisoquinolin-6-yl or tetrahydroisoquinolin-7-yl,each of which is substituted with 0 to 3 substituents independentlychosen from C₁-C₃alkyl.

Compounds and salts of Formula I and Formula II are provided herein inwhich R_(A) is tetrahydroisoquinolin-6-yl substituted at the 1, 2, and 3positions with 0 to 3 methyl substituents.

Further included are compounds and salts of Formula I and Formula II inwhich

R_(A) is isoindol-5-yl substituted with 0 to 3 independently chosenC₁-C₃alkyl substituents.

In still other embodiments the invention provides compounds and salts ofFormula I and Formula II, in which R_(A) is isoindol-5-yl substituted atthe 1, 2, or 3 positions with 0 to 3 methyl substituents.

Still other embodiments of the invention include compounds and salts ofFormula I and Formula II in which R₇ is XR_(B) where X is absent,CH₂—CH₂—, —CH═CH—, or —C═C—; and R_(B) is phenyl; substituted with 1 to5 substituents independently chosen from (i), (ii), and (iii), wherein

-   -   (i) is chosen from halogen, hydroxy, amino, cyano, and nitro,    -   (ii) is chosen from C₁-C₄alkyl, C₂-C₄alkenyl, C₁-C₄alkoxy, mono-        and di-(C₁-C₄)alkylamino, C₁-C₂haloalkyl, C₁-C₂haloalkoxy,        C₃-C₇cycloalkyl(C₀-C₂alkyl), C₃-C₇cycloalkyl(C₀-C₂alkoxy),        phenyl(C₀-C₂alkyl), phenyl(C₀-C₂alkoxy),        pyrrolidinyl(C₀-C₂alkyl), piperidinyl(C₀-C₂alkyl),        piperazinyl(C₀-C₂alkyl), morpholinyl(C₀-C₂alkyl),        thiomorpholinyl(C₀-C₂alkyl), pyridyl, pyrimidinyl, pyrazinyl,        furanyl, benzofuranyl, pyrrolyl, pyrazolyl, imidazolyl, thienyl,        C₁-C₄alkylthio, ═NOR₁₀, —(C₀-C₄alkyl)(C═O)R₁₀,        —(C₀-C₄alkyl)O(C═O)R₁₀, —(C₀-C₄alkyl)(C═O)NR₁₀R₁₁,        —(C₀-C₄alkyl)O(C═O)NR₁₀R₁₁, —(C₀-C₄alkyl) (C═O)OR₁₀,        —(C₀-C₄alkyl)NR₁₀(C═O)R₁₁, —(C₀-C₄alkyl)NR₁₀(C═O)OR₁₁,        —(C₀-C₄alkyl)NR₁₀(C═O)NR₁₁R₁₂,        —(C₀C₄alkyl)NR₁₀(C═O)(C₁-C₄)NR₁₁(C═O)OR₁₂,        —(C₀-C₄alkyl)NR₁₀(C═S)NR₁₁R₁₂, —(C₀-C₄alkyl)NR₁₀NR₁₁R₁₂,        —(C₀-C₄alkyl)N═NR₁₃, —(C₀-C₄alkyl)SO₃R₁₀,        —(C₀-C₄alkyl)(S═O)OR₁₀, —(C₀-C₄alkyl)SO₂R₁₃,        —(C₀-C₄alkyl)SO₂NR₁₀R₁₁, and —(C₀-C₄alkyl)NR₁₀SO₂R₁₃; and    -   (iii) is chosen from —OR_(D), —(C═O)R_(D), —SO₂R_(D), —SO₃R_(D),        —NR ₁₀SO₂R_(D), where R_(D) is C₄alkyl,        C₃-C₇cycloalkyl(C₀-C₂alkyl), pyrrolidinyl(C₀-C₂alkyl),        piperidinyl(C₀-C₂alkyl), piperazinyl(C₀-C₂alkyl),        morpholinyl(C₀-C₂alkyl), thiomorpholinyl(C₀-C₂alkyl),        phenyl(C₀-C₂alkyl), naphthyl(C₀-C₂alkyl), pyridyl(C₀-C₂alkyl),        pyrimidinyl(C₀-C₂alkyl), pyrazinyl(C₀-C₂alkyl),        furanyl(C₀-C₂alkyl), benz[b]thiophenyl(C₀-C₂alkyl),        benzofuranyl(C₀-C₂alkyl), quinolinyl(C₀-C₂alkyl),        isoquinolinyl(C₀-C₂alkyl), quinazolinyl(C₀-C₂alkyl),        isoxazolyl(C₀-C₂alkyl), indolyl(C₀-C₂alkyl),        dihydroindolyl(C₀-C₂alkyl), pyrrolyl(C₀-C₂alkyl),        pyrazolyl(C₀-C₂alkyl), imidazolyl(C₀-C₂alkyl),        thienyl(C₀-C₂alkyl), isoindolyl(C₀-C₂alkyl), and        dihydroisoindolyl(C₀-C₂alkyl).

Each of (ii) and (iii) is substituted with 0 to 3 substitutentsindependently chosen from halogen, hydroxy, amino, cyano, nitro, oxo,—COOH, —CONH₂, C₁-C₄alkyl, C₂-C₄alkenyl, C₂-C₄alkynyl, C₁-C₄alkoxy,C₁-C₄alkoxycarbonyl, C₃-C₇cycloalkyl(C₀-C₄alkyl),C₃-C₇cycloalkyl(C₀-C₄alkoxy), mono- and di-(C₁-C₄)alkylamino,C₁-C₂haloalkyl, C₁-C₂haloalkoxy, C₂-C₄alkanoyl, and phenyl.

The invention also includes compounds and salts of Formula I and FormulaII in which R₇ is XR_(B), where X is absent, CH₂—CH₂—, —CH═CH—, or—C≡C—; R_(B) is phenyl; and

-   -   (i) is chosen from halogen, hydroxy, amino, cyano, and nitro,    -   (ii) is chosen from C₁-C₄alkyl, C₂-C₄alkenyl, C₁-C₄alkoxy, mono-        and di-(C₁-C₄)alkylamino, C₁-C₂haloalkyl, C₁-C₂haloalkoxy,        C₃-C₇cycloalkyl(C₀-C₂alkyl), C₃-C₇cycloalkyl(C₀-C₂alkoxy),        phenyl(C₀-C₂alkyl), phenyl(C₀-C₂alkoxy), pyrrolidinyl,        piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, pyridyl,        thienyl, C₁-C₄alkylthio, —(C═O)OR₁₀, and —(C═O)NR₁₀R₁₁; and    -   (iii) is chosen from —OR_(D), —(C═O)R_(D), —SO₂R_(D), —SO₃R_(D),        and —NR₁₀SO₂R_(D), where R_(D) is C₁-C₄alkyl,        C₃-C₇cycloalkyl(C₀-C₂alkyl), piperidinyl, piperazinyl, phenyl,        naphthyl, and pyridyl.

Each of (ii) and (iii) in this embodiment is substituted with 0 to 3substituents independently chosen from halogen, hydroxy, amino, cyano,nitro, oxo, —COOH, —CONH₂, C₁-C₄alkoxy, C₁-C₄alkoxycarbonyl,C₃-C₇cycloalkyl(C₀-C₂alkyl), mono- and di-(C₁-C₄)alkylamino,C₁-C₂haloalkyl, C₁-C₂haloalkoxy, C₂-C₄alkanoyl, and phenyl.

The invention further includes compounds and salts of Formula I andFormula II in which R₇ is XR_(B) and X is absent, CH₂—CH₂—, —CH═CH—, or—C≡C-. R_(B) is phenyl; substituted with 0 to 3 substituentsindependently chosen from halogen, hydroxy, amino, cyano, nitro,C₁-C₄alkyl, C₂-C₄alkenyl, C₁-C₄alkoxy, mono- and di-(C₁-C₄)alkylamino,C₁-C₂haloalkyl, C₁-C₂haloalkoxy, C₃-C₇cycloalkyl(C₀-C₂alkyl),C₃-C₇cycloalkyl(C₀-C₂alkoxy), phenyl(C₀-C₂alkyl), phenyl(C₀-C₂alkoxy),pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl,pyridyl, thienyl, C₁-C₄alkylthio, —(C═O)OR₁₀, and —(C═O)NR₁₀R₁₁; and

R_(B) is substituted with 1 or 2 substituents independently chosen from—OR_(D), —(C═O)R_(D), —SO₂R_(D), —SO₃R_(D), and —NR₁₀SO₂R_(D), whereR_(D) is C₁-C₄alkyl, piperidinyl, phenyl, naphthyl, or pyridyl; and eachR_(D) is substituted with 0 to 3 substituents independently chosen fromhalogen, hydroxy, amino, cyano, C₁-C₂alkyl, C₁-C₂alkoxy, mono- anddi-(C₁-C₂)alkylamino, C₁-C₂haloalkyl, and C₁-C₂haloalkoxy.

The invention includes compounds and salts of Formula I and Formula IIin which R₇ is XR_(B) and X is absent.

The invention pertains to certain compounds of Formula I and Formula IIin which R₇ is XR_(B) and R_(B) is phenyl substituted with 1 to 3substituents independently chosen from halogen, hydroxy, amino, cyano,C₁-C₄alkyl, C₁-C₄alkoxy, mono- and di-(C₁-C₄)alkylamino, C₁-C₂haloalkyl,and C₁-C₂haloalkoxy.

Certain embodiments of the invention also pertain to compounds and saltsof Formula I and Formula II in which R₇ is XR_(B) and R_(B) is phenyl,substituted with 1 or 2 substituents independently chosen from halogen,hydroxy, amino, C₁-C₂alkyl, and C₁-C₂alkoxy.

Other embodiments of the invention pertain to compounds and salts ofFormula I and Formula II in which phenyl substituted with 1 or 2substituents independently chosen from fluoro, amino, hydroxy, cyano,and methyl.

In other embodiments the invention provides compounds and salts ofFormula I and Formula II in which

R₇ is XR_(D) where R_(D) is phenyl fused to a 5- or 6-memberedheterocycloalkyl ring containing 1 or 2 nitrogen or oxygen atoms, whereR_(D) is substituted with 0 to 3 substitutents independently chosen from(i), (ii), and (iii).

-   -   (i) is chosen from halogen, hydroxy, amino, cyano, and nitro,    -   (ii) is chosen from C₁-C₄alkyl, C₂-C₄alkenyl, C₁-C₄alkoxy, mono-        and di-(C₁-C₄)alkylamino, C₁-C₂haloalkyl, C₁-C₂haloalkoxy,        C₃-C₇cycloalkyl(C₀-C₂alkyl), C₃-C₇cycloalkyl(C₀-C₂alkoxy),        phenyl(C₀-C₂alkyl), phenyl(C₀-C₂alkoxy),        pyrrolidinyl(C₀-C₂alkyl), piperidinyl(C₀-C₂alkyl),        piperazinyl(C₀-C₂alkyl), morpholinyl(C₀-C₂alkyl),        thiomorpholinyl(C₀-C₂alkyl), pyridyl, pyrimidinyl, pyrazinyl,        furanyl, benzofuranyl, pyrrolyl, pyrazolyl, imidazolyl, thienyl,        C₁-C₄alkylthio, ═NOR₁₀, —(C₀-C₄alkyl)(C═O)R₁₀,        —(C₀-C₄alkyl)O(C═O)R₁₀, —(C₀-C₄alkyl)(C═O)NR₁₀R₁₁,        —(C₀-C₄alkyl)O(C═O)NR₁₀R₁₁, —(C₀-C₄alkyl) (C═O)OR₁₀,        —(C₀-C₄alkyl)NR₁₀(C═O)R₁₁, —(C₀-C₄alkyl)NR₁₀(C═O)OR₁₁,        —(C₀-C₄alkyl)NR₁₀(C═O)NR₁₁R₁₂,        —(C₀-C₄alkyl)NR₁₀(C═O)(C₁-C₄alkyl)NR₁₁(C═O)O—R₁₂,        —(C₀-Colkyl)NR₁₀(C═S)NR₁₁R₁₂, —(C₀-C₄alkyl)NR₁₀NR₁₁R₁₂,        —(C₀-C₄alkyl)N═NR₁₃, —(C₀-C₄alkyl)SO₃R₁₀,        —(C₀-C₄alkyl)(S═O)OR₁₀, —(C₀-C₄alkyl)SO₂R₁₃,        —(C₀-C₄alkyl)SO₂NR₁₀R₁₁, and —(C₀-C₄alkyl)NR₁₀SO₂R₁₃; and    -   (iii) is chosen from —OR_(D), —(C═O)R_(D), —SO₂R_(D), —SO₃R_(D),        —NR₁₀SO₂R_(D), where R_(D) is C₁-C₄alkyl,        C₃-C₇cycloalkyl(C₀-C₂alkyl), pyrrolidinyl(C₀-C₂alkyl),        piperidinyl(C₀-C₂alkyl), piperazinyl(C₀-C₂alkyl),        morpholinyl(C₀-C₂alkyl), thiomorpholinyl(C₀-C₂alkyl),        phenyl(C₀-C₂alkyl), naphthyl(C₀-C₂alkyl), pyridyl(C₀-C₂alkyl),        pyrimidinyl(C₀-C₂alkyl), pyrazinyl(C₀-C₂alkyl),        furanyl(C₀-C₂alkyl), benz[b]thiophenyl(C₀-C₂alkyl),        benzofuranyl(C₀-C₂alkyl), quinolinyl(C₀-C₂alkyl),        isoquinolinyl(C₀-C₂alkyl), quinazolinyl(C₀-C₂alkyl),        isoxazolyl(C₀-C₂alkyl), indolyl(C₀-C₂alkyl),        dihydroindolyl(C₀-C₂alkyl), pyrrolyl(C₀-C₂alkyl),        pyrazolyl(C₀-C₂alkyl), imidazolyl(C₀-C₂alkyl),        thienyl(C₀-C₂alkyl), isoindolyl(C₀-C₂alkyl), or        dihydroisoindolyl(C₀-C₂alkyl); where each of (ii) and (iii) is        substituted with 0 to 3 substitutents independently chosen from        halogen, hydroxy, amino, cyano, nitro, oxo, —COOH, —CONH₂,        C₁-C₄alkyl, C₂-C₄alkenyl, C₂-C₄alkynyl, C₁-C₄alkoxy,        C₁-C₄alkoxycarbonyl, C₃-C₇cycloalkyl(C₀-C₄alkyl),        C₃-C₇cycloalkyl(C₀-C₄alkoxy), mono- and di-(C₁-C₄)alkylamino,        C₁-C₂haloalkyl, C₁-C₂haloalkoxy, C₂-C₄alkanoyl, and phenyl.

In certain embodiments provided herein R₇ is XR_(D) and X is absent.

In certain embodiments provided herein R₇ is XR_(D) and X is absent andR_(D) is phenyl fused to a 5- or 6-membered heterocycloalkyl ringcontaining 1 or 2 nitrogen or oxygen atoms, where R_(D) is substitutedwith 0 to 2 substitutents independently chosen from halogen, hydroxy,amino, C₁-C₂alkyl, and C₁-C₂alkoxy.

The A₈ Variable

The invention includes compounds and salts of Formula I and Formula IIin which A₈ is nitrogen. Examples of such compounds include, but are notlimited to, compounds of Formula III-Formula VI

The invention also includes compounds and salts of Formula I and FormulaII in which A₈ is CR₈. Examples of such compounds include, but are notlimited to, compounds of Formula VII-Formula X

The variables R₂, R₃, R₄, R₅, R₆, R₇, R₈, and R₉ shown in FormulaIII-Formula X carry any of the definitions set forth herein for thesevariables.

The invention includes compounds and salts of Formula I and Formula IIin which R₈ is hydrogen, halogen, C₁-C₂alkyl, C₁-C₂alkoxy,C₁-C₂haloalkyl, or C₁-C₂haloalkoxy.

Certain embodiments of the invention pertain to compounds and salts ofFormula I and Formula II in which R₈ is hydrogen or methoxy.

The R₉ Variable

The invention includes compounds and salts of Formula I and Formula IIin which R₉ is C₁-C₄alkyl, cyclopropyl, or phenyl, each of which issubstituted with 0 to 3 substituents independently chosen from halogen,hydroxy, amino, C₁-C₂alkyl, C₁-C₂alkoxy, mono- and di-(C₁-C₂)alkylamino,C₁-C₂haloalkyl, and C₁-C₂haloalkoxy.

Other embodiments of the invention pertain to compounds and salts ofFormula I and Formula II in which R₉ is C₁-C₄alkyl or cyclopropyl, or R₉is phenyl substituted with 2 substituents chosen from halogen, hydroxy,amino, C₁-C₂alkyl, C₁-C₂alkoxy, mono- and di-(C₁-C₂)alkylamino,C₁-C₂haloalkyl, and C₁-C₂haloalkoxy.

Certain embodiments of the invention include compounds and salts ofFormula I and Formula II in which R₉ is ethyl, t-butyl, cyclopropyl or2,4-difluorophenyl, and particularly include those compounds and saltsin which R₉ is cyclopropyl.

The invention includes compounds of Formula I and Formula II in whichthe variables A₁, R₂, R₃, R₅, R₆, R₇, A₈ and R₉ carry any combination ofthe definitions set forth for these variables above.

Certain compounds of Formula I and Formula II exhibit possess potentantibacterial, antifungal, and/or antiprotozoal activity. Particularcompounds of the invention exhibit Minimum Inhibitory Concentrations(MIC) of 64 μg/ml or less against Staphyloccocus aureus and/orEschericia coli in a standard assay for determining the MIC of acompound against these bacteria, such as the assay provided in Example10 below. Preferred compounds of the Formula I and II exhibit MIC valuesof 10 μg/ml or less against Staphyloccocus aureus and/or Eschericiacoli. More preferred compound of the Formula I and II exhibit MIC valuesof 4 μg/ml or less, or even more preferably 1 μg/ml or less, againstStaphyloccocus aureus and/or Eschericia coli.

Certain compounds of Formula I and Formula II are selectiveantimicrobial agents; having the ability to kill or inhibit the growthor reproduction of microbial organisms, while having little or no effecton the cells of fish, amphibians, reptiles, birds, or mammals. Theselectivity of compounds of Formula I and Formula II may be assessed bydetermining the CC₅₀ (the concentration at which 50% of the cells arekilled) for cultured cells of a higher animal, such as a fish, reptiles,amphibian, bird, or mammal. Certain compounds of the invention exhibit aCC₅₀ of greater that 100 micromolar for mammalian cells. Certaincompounds of the invention exhibit a CC₅₀ of greater than 100 micromolarfor cultured human hepatocytes, and also exhibit MIC values of 64 μg/mlor less, preferably 10 μg/ml or less, or more preferably 4 μg/ml orless, or still more preferably 1 μg/ml or less against Staphyloccocusaureus and/or Eschericia coli.

Without wishing to be bound to any particular theory it is believed thatthe antimicrobial properties of compounds of Formula I and Formula IIare due to the ability to these compounds to inhibit the activity ofmicrobial DNA gyrases while having little or no effect on the analogousenzyme, Topoisomerase II, present in higher organisms. Certain preferredcompounds of the invention are 100-fold or more selective for bacterialDNA gyrases than for mammalian, particularly human, Topoisomerase II.

Synthetic Intermediates

The invention includes novel intermediates useful for the synthesis ofantimicrobial compounds of Formula I and Formula II. Coupling reactionsoccur between R′—M and R″—Y in the presence of catalyst Q, where M isLi, Mg, B, Al, Si, Zn, Cu, Zr, or Sn; where Y is I, Br, Cl, —O(SO₂)CF₃,or —N₂BF₄; Q is Fe, Ni, Cu, Pd, or Rh, and R′ and R″ are the organicmolecules to which M and Y are bound. In certain embodiments M is Boron,disubstituted with OH, OG, or G, where G is an optionally substitutedstraight, branched or cyclic alkyl group, or other suitable group; Y isBr, and where Q is Pd. A general review of this chemistry can be foundin Tamao, K. and Miyaura, N. Topics in Current Chemistry 219: 1-9(2002). A review of the use of coupling reagents in which M is Boronwith a listing of potential boronates, palladium catalysts, and reactionconditions can be found in Miyaura, N. Topics in Current Chemistry 219:11-59 (2002).

Thus the invention includes intermediates of Formula XI and Formula XII:

In which A₁, A₈, R₂, R₃, R₅, R₆, and R₉ carry the definitions set forthabove and Y is I, Br, Cl, —O(SO₂)CF₃, or —N₂BF₄. Here compounds ofFormula XI and Formula XII play the role of R″—Y intermediate in thecoupling reaction between R′—M and R″—Y. These intermediates are coupledto compounds of the Formula R′—M where R′ is a group R.

R is XR_(A) where X is absent, CH₂—CH₂—, —CH═CH—, or —C≡C—, and R_(A) isC₃-C₆alkyl, C₄-C₇cycloalkyl, C₄-C₇cycloalkenyl, a 5-6 memberedsaturated, partially unsaturated, or aromatic heterocylic group boundvia a carbon atom when X is absent or —CH₂—CH₂—, or bound via a carbonor nitrogen atom when X is —CH═CH— or —C≡C— or a R_(A) is a 7-10membered bicyclic saturated, partially unsaturated, or aromaticheterocylic group bound via a carbon atom when X is absent or —CH₂—CH₂—,or bound via a carbon or nitrogen atom when X is —CH═CH— or —C≡C—; eachof which R_(A) is substituted with 0 to 5 substituents independentlychosen from (i), (ii), and (iii); or

R is XR_(B), where R_(B) is phenyl substituted with 1 to 5 substituentsindependently chosen from (i), (ii), and (iii); or

R is XR_(C), where R_(C) is cyclopropyl with 0 to 5 substituentsindependently chosen from (i), (ii), and (iii), with the proviso thatR_(c) is not substituted with amino, or mono- or di-(C₁-C₄)alkylamino.

Wherein (i) is chosen from halogen, hydroxy, amino, cyano, and nitro,

-   -   (ii) is chosen from C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl,        mono- and di-(C₁-C₄)alkylamino, C₁-C₂haloalkyl, C₁-C₂haloalkoxy,        C₃-C₇cycloalkyl(C₀-C₄alkyl), C₃-C₇cycloalkyl(C₀-C₄alkoxy),        C₄-C₇cycloalkenyl(C₀-C₄alkyl), aryl(C₀-C₆carbohydryl),        aryl(C₁-C₄alkoxy), C₂-C₆heterocycloalkyl(C₀-C₄alkyl),        heteroaryl(C₀-C₆carbohydryl), C₁-C₆alkylthio,        -(C₀-C₄alkyl)O(C═O)R₁₀, —(C₀-C₄alkyl)(C═O)NR₁₀R₁₁,        —(C₀-C₄alkyl)O(C═O)NR₁₀R₁₁, —(C₀-C₄alkyl) (C═O)OR₁₀,        —(C₀-C₄alkyl)NR₁₀(C═O)R₁₁, —(C₀-C₄alkyl)NR₁₀(C═O)OR₁₁,        —(C₀-C₄alkyl)NR₁₀(C═O)NR₁₁R₁₂, —(C₀-C₄alkyl)NR₁₀(C═S)NR₁₁R₁₂,        —(C₀-C₄alkyl)NR₁₀NR₁₁R₁₂, —(C₀-C₄alkyl)N═NR₁₃,        —(C₀-C₄alkyl)SO₃R₁₀, —(C₀-C₄alkyl)(S═O)OR₁₀,        —(C₀-C₄alkyl)SO₂R₁₃, —(C₀-C₄alkyl)SO₂NR₁₀R₁₁, and        —(C₀-C₄alkyl)NR₁₀SO₂R₁₃; and    -   (iii) is chosen from —OR_(D), —(C═O)R_(D), —SO₂R_(D), —SO₃R_(D),        —NR₁₀SO₂R_(D), where R_(D) is C₁-C₄alkyl,        C₃-C₇cycloalkyl(C₀-C₂alkyl), C₂-C₆heterocycloalkyl(C₀-C₂alkyl),        aryl(C₀-C₂alkyl), or heteroaryl(C₀-C₂alkyl); where each of (ii)        and (iii) is substituted with 0 to 3 substituents independently        chosen from halogen, hydroxy, amino, cyano, nitro, —COOH,        —(C═O)OCH₃, —CONH₂, C₁-C₄alkyl, C₂-C₄alkenyl, C₂-C₄alkynyl,        C₁-C₄alkoxy, C₃-C₇cycloalkyl(C₀-C₄alkyl),        C₃-C₇cycloalkyl(C₀-C₄alkoxy), mono- and di-(C₁-C₄)alkylamino,        C₁-C₂haloalkyl, C₁-C₂haloalkoxy, and C₂-C₄alkanoyl.

M is Li, Mg, B, Al, Si, Zn, Cu, Zr, or Sn; or M is Boron, disubstitutedwith OH, OG, or G, where G is an optionally substituted straight,branched, or cyclic alkyl group, an optionally substituted aryl orarylalkyl group, or other suitable group.

The invention also includes intermediates of Formula XIII and FormulaXIV in which

In which A₁, A₈, R₂, R₃, R₅, R₆, and R₉\ carry the definitions set forthabove and M Li, Mg, B, Al, Si, Zn, Cu, Zr, or Sn; or M is Boron,disubstituted with OH, OG, or G, where G is an optionally substitutedstraight, branched, or cyclic alkyl group, an optionally substitutedaryl or arylalkyl group, or other suitable group. Here compounds ofFormula XIII and Formula XIV play the role of R′—M in the couplingreaction. These intermediates are coupled to compounds of the FormulaR″—Y where Y is I, Br, Cl, —O(SO₂)CF₃, or —N₂BF₄ and R″ carries thedefinition set forth above for R.

Pharmaceutical Preparations

Compounds and salts of Formula I and Formula II can be administered asthe neat chemical, but are preferably administered as a pharmaceuticalcomposition or formulation. Accordingly, the invention providespharmaceutical formulations comprising a compound or pharmaceuticallyacceptable salt of Formula I or Formula II, together with one or morepharmaceutically acceptable carrier, excipients, adjuvant, diluent,excipient, or other ingredient.

Compounds of general Formula I and Formula II may be administeredorally, topically, parenterally, by inhalation or spray, sublingually,transdermally, via buccal administration, rectally, as an ophthalmicsolution, or by other means, in dosage unit formulations containingconventional non-toxic pharmaceutically acceptable carriers, excipients,adjuvants, and vehicles.

A pharmaceutical composition comprising a compound or salt of Formula Ior Formula II wherein the composition is formulated as an injectablefluid, an aerosol, a cream, a gel, a pill, a capsule, a tablet, a syrup,a transdermal patch, or an ophthalmic solution is provided herein.

In addition to the subject compound, the compositions of the inventionmay contain a pharmaceutically acceptable carrier, one or morecompatible solid or liquid filler diluents or encapsulating substances,which are suitable for administration to an animal. Carriers must be ofsufficiently high purity and sufficiently low toxicity to render themsuitable for administration to the animal being treated. The carrier canbe inert or it can possess pharmaceutical benefits of its own. Theamount of carrier employed in conjunction with the compound issufficient to provide a practical quantity of material foradministration per unit dose of the compound.

Exemplary pharmaceutically acceptable carriers or components thereof aresugars, such as lactose, glucose and sucrose; starches, such as cornstarch and potato starch; cellulose and its derivatives, such as sodiumcarboxymethyl cellulose, ethyl cellulose, and methyl cellulose; powderedtragacanth; malt; gelatin; talc; solid lubricants, such as stearic acidand magnesium stearate; calcium sulfate; vegetable oils, such as peanutoil, cottonseed oil, sesame oil, olive oil, and corn oil; polyols suchas propylene glycol, glycerine, sorbitol, mannitol, and polyethyleneglycol; alginic acid; emulsifiers, such as the TWEENS; wetting agents,such sodium lauryl sulfate; coloring agents; flavoring agents; tabletingagents, stabilizers; antioxidants; preservatives; pyrogen-free water;isotonic saline; and phosphate buffer solutions.

In particular, pharmaceutically acceptable carriers for systemicadministration include sugars, starches, cellulose and its derivatives,malt, gelatin, talc, calcium sulfate, vegetable oils, synthetic oils,polyols, alginic acid, phosphate buffer solutions, emulsifiers, isotonicsaline, and pyrogen-free water. Preferred carriers for parenteraladministration include propylene glycol, ethyl oleate, pyrrolidone,ethanol, and sesame oil.

Optional active agents may be included in a pharmaceutical composition,which do not substantially interfere with the activity of the compoundof the present invention.

Effective concentrations of one or more of the compounds of theinvention including pharmaceutically acceptable salts, esters or otherderivatives thereof, are mixed with one or more suitable pharmaceuticalcarrier, excipient, adjuvant, or vehicle. In instances in which thecompounds exhibit insufficient solubility, methods for solubilizingcompounds may be used. Such methods are known to those of skill in thisart, and include, but are not limited to using cosolvents, such asdimethylsulfoxide (DMSO), using surfactants, such as Tween, ordissolution in aqueous sodium bicarbonate. Derivatives of the compounds,such as salts of the compounds or prodrugs of the compounds may also beused in formulating effective pharmaceutical compositions.

Upon mixing or addition of the compound(s) of Formula I and/or FormulaII, the resulting mixture may be a solution, suspension, emulsion or thelike. The form of the resulting mixture depends upon a number offactors, including the intended mode of administration and thesolubility of the compound in the chosen carrier or vehicle. Theeffective concentration sufficient for ameliorating the symptoms of thedisease, disorder or condition treated and may be empiricallydetermined.

The pharmaceutical compositions containing compounds of general FormulaI and/or Formula II may be in a form suitable for oral use, for example,as tablets, troches, lozenges, aqueous or oily suspensions, dispersiblepowders or granules, emulsions, hard or soft capsules, or syrups orelixirs. Compositions intended for oral use may be prepared according toany method known to the art for the manufacture of pharmaceuticalcompositions and such compositions may contain one or more agents, suchas sweetening agents, flavoring agents, coloring agents and preservingagents, in order to provide pharmaceutically elegant and palatablepreparations.

Oral formulations contain between 0.1 and 99% of a compound of theinvention and usually at least about 5% (weight %) of a compound of thepresent invention. Some embodiments contain from about 25% to about 50%or from 5% to 75% of a compound of invention.

Liquids Formulations

Compounds of the invention can be incorporated into oral liquidpreparations such as aqueous or oily suspensions, solutions, emulsions,syrups, or elixirs, for example. Moreover, formulations containing thesecompounds can be presented as a dry product for constitution with wateror other suitable vehicle before use. Such liquid preparations cancontain conventional additives, such as suspending agents (e.g.,sorbitol syrup, methyl cellulose, glucose/sugar, syrup, gelatin,hydroxyethyl cellulose, carboxymethyl cellulose, aluminum stearate gel,and hydrogenated edible fats), emulsifying agents (e.g., lecithin,sorbitan monsoleate, or acacia), non-aqueous vehicles, which can includeedible oils (e.g., almond oil, fractionated coconut oil, silyl esters,propylene glycol and ethyl alcohol), and preservatives (e.g., methyl orpropyl p-hydroxybenzoate and sorbic acid).

Orally administered compositions also include liquid solutions,emulsions, suspensions, powders, granules, elixirs, tinctures, syrups,and the like. The pharmaceutically acceptable carriers suitable forpreparation of such compositions are well known in the art. Oralformulations may contain preservatives, flavoring agents, sweeteningagents, such as sucrose or saccharin, taste-masking agents, and coloringagents.

Typical components of carriers for syrups, elixirs, emulsions andsuspensions include ethanol, glycerol, propylene glycol, polyethyleneglycol, liquid sucrose, sorbitol and water. Syrups and elixirs may beformulated with sweetening agents, for example glycerol, propyleneglycol, sorbitol or sucrose. Such formulations may also contain ademulcent.

Suspensions

For a suspension, typical suspending agents include methylcellulose,sodium carboxymethyl cellulose, AVICEL RC-591, tragacanth and sodiumalginate; typical wetting agents include lecithin and polysorbate 80;and typical preservatives include methyl paraben and sodium benzoate.

Aqueous suspensions contain the active material(s) in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydropropylmethylcellulose,sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents; naturally-occurring phosphatide, forexample, lecithin, or condensation products of an alkylene oxide withfatty acids, for example polyoxyethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethyleneoxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol substitute, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides, for example polyethylene sorbitan substitute.The aqueous suspensions may also contain one or more preservatives, forexample ethyl, or n- propyl p-hydroxybenzoate.

Oily suspensions may be formulated by suspending the active ingredientsin a vegetable oil, for example peanut oil, olive oil, sesame oil orcoconut oil, or in a mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and flavoring agents may be added to provide palatable oralpreparations. These compositions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

Emulsions

Pharmaceutical compositions of the invention may also be in the form ofoil-in-water emulsions. The oily phase may be a vegetable oil, forexample olive oil or peanut oil, or a mineral oil, for example liquidparaffin or mixtures of these. Suitable emulsifying agents may benaturally-occurring gums, for example gum acacia or gum tragacanth,naturally-occurring phosphatides, for example soy bean, lecithin, andesters or partial esters derived from fatty acids and hexitol,anhydrides, for example sorbitan monoleate, and condensation products ofthe said partial esters with ethylene oxide, for example polyoxyethylenesorbitan monoleate.

Dispersible Powders

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.

Tablets and Capsules

Tablets typically comprise conventional pharmaceutically compatibleadjuvants as inert diluents, such as calcium carbonate, sodiumcarbonate, mannitol, lactose and cellulose; binders such as starch,gelatin and sucrose; disintegrants such as starch, alginic acid andcroscarmelose; lubricants such as magnesium stearate, stearic acid andtalc. Glidants such as silicon dioxide can be used to improve flowcharacteristics of the powder mixture. Coloring agents, such as the FD&Cdyes, can be added for appearance. Sweeteners and flavoring agents, suchas aspartame, saccharin, menthol, peppermint, and fruit flavors, areuseful adjuvants for chewable tablets. Capsules (including time releaseand sustained release formulations) typically comprise one or more soliddiluents disclosed above. The selection of carrier components oftendepends on secondary considerations like taste, cost, and shelfstability.

Such compositions may also be coated by conventional methods, typicallywith pH or time-dependent coatings, such that the subject compound isreleased in the gastrointestinal tract in the vicinity of the desiredtopical application, or at various times to extend the desired action.Such dosage forms typically include, but are not limited to, one or moreof cellulose acetate phthalate, polyvinylacetate phthalate,hydroxypropyl methylcellulose phthalate, ethyl cellulose, Eudragitcoatings, waxes and shellac.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, for example peanut oil, liquid paraffin or olive oil.

Injectable and Parenteral Formulations

Pharmaceutical compositions may be in the form of a sterile injectableaqueous or oleaginous suspension. This suspension may be formulatedaccording to the known art using those suitable dispersing or wettingagents and suspending agents that have been mentioned above. The sterileinjectable preparation may also be sterile injectable solution orsuspension in a non-toxic parentally acceptable diluent or solvent, forexample as a solution in 1,3-butanediol. Acceptable vehicles andsolvents include water, Ringer's solution, and isotonic sodium chloridesolution. In addition, sterile, fixed oils are conventionally employedas a solvent or suspending medium. For this purpose any bland fixed oilmay be employed including synthetic mono- or diglycerides. In addition,fatty acids such as oleic acid are useful in the preparation ofinjectables.

Compounds of Formula I and Formula II may be administered parenterallyin a sterile medium. Parenteral administration includes subcutaneousinjections, intravenous, intramuscular, intrathecal injection, orinfusion techniques. The drug, depending on the vehicle andconcentration used, can either be suspended or dissolved in the vehicle.Advantageously, adjuvants such as local anesthetics, preservatives andbuffering agents can be dissolved in the vehicle. In compositions forparenteral administration the carrier comprises at least about 90% byweight of the total composition.

Suppositories

Compounds of Formula I and Formula II may also be administered in theform of suppositories for rectal administration of the drug. Thesecompositions can be prepared by mixing the drug with a suitablenon-irritating excipient that is solid at ordinary temperatures butliquid at the rectal temperature and will therefore melt in the rectumto release the drug. Such materials are cocoa butter and polyethyleneglycols.

Topical Formulations

Compounds of the invention may be formulated for local or topicalapplication, such as for topical application to the skin and mucousmembranes, such as in the eye, in the form of gels, creams, and lotionsand for application to the eye or for intracisternal or intraspinalapplication. Topical compositions of the present invention may be in anyform including, for example, solutions, creams, ointments, gels,lotions, milks, cleansers, moisturizers, sprays, skin patches, and thelike.

Such solutions may be formulated as 0.01% -10% isotonic solutions, pHabout 5-7, with appropriate salts. Compounds of the invention may alsobe formulated for transdermal administration as a transdermal patch.

Topical compositions containing the active compound can be admixed witha variety of carrier materials well known in the art, such as, forexample, water, alcohols, aloe vera gel, allantoin, glycerine, vitamin Aand E oils, mineral oil, propylene glycol, PPG-2 myristyl propionate,and the like.

Other materials suitable for use in topical carriers include, forexample, emollients, solvents, humectants, thickeners and powders.Examples of each of these types of materials, which can be used singlyor as mixtures of one or more materials, are as follows:

Emollients, such as stearyl alcohol, glyceryl monoricinoleate, glycerylmonostearate, propane-1,2-diol, butane-1,3-diol, mink oil, cetylalcohol, iso-propyl isostearate, stearic acid, iso-butyl palmitate,isocetyl stearate, ( )ey' alcohol, isopropyl laurate, hexyl laurate,decyl oleate, octadecan-2-ol, isocetyl alcohol, cetyl palmitate,dimethylpolysiloxane, di-n-butyl sebacate, iso-propyl myristate,iso-propyl palmitate, iso-propyl stearate, butyl stearate, polyethyleneglycol, triethylene glycol, lanolin, sesame oil, coconut oil, arachisoil, castor oil, acetylated lanolin alcohols, petroleum, mineral oil,butyl myristate, isostearic acid, palmitic acid, isopropyl linoleate,lauryl lactate, myristyl lactate, decyl oleate, and myristyl myristate;propellants, such as propane, butane, iso-butane, dimethyl ether, carbondioxide, and nitrous oxide; solvents, such as ethyl alcohol, methylenechloride, iso-propanol, castor oil, ethylene glycol monoethyl ether,diethylene glycol monobutyl ether, diethylene glycol monoethyl ether,dimethyl sulphoxide, dimethyl formamide, tetrahydrofuran; humectants,such as glycerin, sorbitol, sodium 2-pyrrolidone-5-carboxylate, solublecollagen, dibutyl phthalate, and gelatin; and powders, such as chalk,talc, fullers earth, kaolin, starch, gums, colloidal silicon dioxide,sodium polyacrylate, tetra alkyl ammonium smectites, trialkyl arylammonium smectites, chemically modified magnesium aluminium silicate,organically modified montmorillonite clay, hydrated aluminium silicate,fumed silica, carboxyvinyl polymer, sodium carboxymethyl cellulose, andethylene glycol monostearate.

The compounds of the invention may also be topically administered in theform of liposome delivery systems, such as small unilamellar vesicles,large unilamellar vesicles, and multilamellar vesicles. Liposomes can beformed from a variety of phospholipids, such as cholesterol,stearylamine or phosphatidylcholines.

Other Formulations

Other compositions useful for attaining systemic delivery of the subjectcompounds include sublingual, buccal and nasal dosage forms. Suchcompositions typically comprise one or more of soluble filler substancessuch as sucrose, sorbitol and mannitol, and binders such as acacia,microcrystalline cellulose, carboxymethyl cellulose and hydroxypropylmethylcellulose. Glidants, lubricants, sweeteners, colorants,antioxidants and flavoring agents disclosed above may also be included.

Compositions for inhalation typically can be provided in the form of asolution, suspension or emulsion that can be administered as a drypowder or in the form of an aerosol using a conventional propellant(e.g., dichlorodifluoromethane or trichlorofluoromethane).

Additional Components

The compositions of the present invention may also optionally comprisean activity enhancer. The activity enhancer can be chosen from a widevariety of molecules that function in different ways to enhanceantimicrobial effects of compounds of the present invention. Particularclasses of activity enhancers include skin penetration enhancers andabsorption enhancers.

Pharmaceutical compositions of the invention may also contain additionalactive agents chosen from a wide variety of molecules, which canfunction in different ways to enhance the antimicrobial or therapeuticeffects of a compound of the present invention. These optional otheractive agents, when present, are typically employed in the compositionsof the invention at a level ranging from about 0.01% to about 15%. Someembodiments contain from about 0.1% to about 10% by weight of thecomposition. Other embodiments contain from about 0.5% to about 5% byweight of the composition.

Packaged Formulations

The invention includes packaged pharmaceutical formulations. Suchpackaged formulations include a pharmaceutical composition containingone or more compounds or salts of Formula I or Formula II in a containerand optionally include instructions for using the composition to treatan animal (typically a human patient) suffering from a microorganisminfection or prevent a microorganism infection in an animal. In certainembodiments the instructions are instructions for using the compositionto treat a patient suffering from a bacterial infection.

In all of the foregoing embodiments the compound of the invention can beadministered alone or as mixtures, and the compositions may furtherinclude additional drugs or excipients as appropriate for theindication.

Methods of Treatment

The invention includes methods of preventing and treating microorganisminfections, particularly bacterial and protozoal infections, byadministering a therapeutically effective amount of one or morecompounds of Formula I and of Formula II to an animal at risk for amicroorganism infection or suffering from a microorganism infection. Theanimal may be a fish, amphibian, reptile or bird, but is preferably amammal. Methods of treating and preventing microorganism infections inlivestock animals, companion animals, and human patients areparticularly preferred.

The compounds disclosed herein are useful for preventing and treatingbacterial infections in animals. Furthermore compounds of the inventionmay be used to treat a variety of conditions not attributed to bacterialinfections. These include diseases and disorders caused fungalinfections, mycoplasma infections, protozoal infections, or otherconditions involving infectious organisms.

In some circumstances an effective amount of a compound of Formula I orFormula II may be an amount sufficient to reduce the symptoms of themicroorganism infection. Alternatively an effective amount of a Compoundof Formula I may be an amount sufficient to significantly reduce theamount of microorganism or antibodies against the detectable in apatient's tissues or bodily fluids.

Methods of treatment also include inhibiting microorganism replicationin vivo, in an animal at risk for a microorganism infection or sufferingfrom such an infection, by administering a sufficient concentration of acompound of Formula I or Formula II to inhibit bacterial survival invitro. By “sufficient concentration” of a compound administered to thepatient is meant the concentration of the compound available in theanimal's system to prevent or combat the infection. Such a concentrationby be ascertained experimentally, for example by assaying bloodconcentration of the compound, or theoretically, by calculatingbioavailability. The amount of a compound sufficient to inhibitbacterial survival in vitro may be determined with a conventional assayfor bacterial survival such as the Minimum Inhibitory Concentration(MIC) Assay disclosed in Example 10, which follows.

The invention also includes using compounds of Formula I and Formula Iin prophylactic therapies. In the context of prophylactic orpreventative treatment an effective amount of a compound of theinvention is an amount sufficient to significantly decrease the treatedanimal's risk of contracting a microorganism infection.

Compounds of the invention are particularly useful for treating andpreventing infectious disorders. These include for example: ocularinfections such as conjunctivitis; urinary tract and genital infections,such as complicated urinary tract infections, acute urinary tract andgenital infections, such as pyelonephritis, cervical gonococcalinfections, cystitis, urethral chlamydial infections, cervicalchlamydial infections, urethral gonococcal infections, and prostatitis,respiratory infections, such as lower respiratory tract infections,acute sinusitis, acute exacerbations of chronic bronchitis,community-acquired pneumonia, and nosocomial pneumonia, skin infections,such as skin-structure infections, impetigo, folliculitis, boils,scalded skin syndrome, and cellulites, and other infections such as boneinfections, joint infections, infectious diarrhea, typhoid fever,intra-abdominal infections, gynecologic infections, including toxicshock syndrome, pelvic infections, and post-surgical infections.

The disclosed compounds are useful for treating infections caused by thefollowing microorganisms:

Aerobic Gram-positive Microorganisms: Including but not limited toEnterococcus faecalis, Enterococcus faecium, Staphylococcus aureus,Staphylococcus epidermidis, Staphylococcus saprophyticus, Streptococcuspneumoniae, Streptococcus pyogenes, Staphylococcus haemolyticus, andStaphylococcus hominis.

Aerobic Gram-negative Microorganisms: Including but not limited toCampylobacter jejuni, Citrobacter diversus, Citrobacter freundii,Enterobacter cloacae, Escherichia coli, Haemophilus influenzae,Haemophilus parainfluenzae, Klebsiella pneumoniae, Moraxellacatarrhalis, Morganella morganii, Neisseria gonorrhoeae, Proteusmirabilis, Proteus vulgaris, Providencia rettgeri, Providencia stuartii,Pseudomonas aeruginosa, Stenotrophomonas maltophila, Salmonella typhi,Serratia marcescens, Shigella boydii, Shigella dysenteriae, Shigellaflexneri, Shigella sonnei. Acinetobacter Iwoffi, Aeromonas hydrophila,Edwardsiella tarda, Enterobacter aerogenes, Klebsiella oxytoca,Legionella pneumophila, Pasteurella multocida, Salmonella enteritidis,Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus, Yersiniaenterocolitica, and H. Pylorii.

Non-bacterial microorganisms: Mycoplasma, Legionella and Chlamydia.

Dosage levels of the order of from about 0.1 mg to about 140 mg perkilogram of body weight per day are useful in the treatment of theabove-indicated conditions (about 0.5 mg to about 7 g per patient perday). The amount of active ingredient that may be combined with thecarrier materials to produce a single dosage form will vary dependingupon the host treated and the particular mode of administration. Dosageunit forms will generally contain between from about 1 mg to about 500mg of an active ingredient.

Frequency of dosage may also vary depending on the compound used and theparticular disease treated. However, for treatment of most infectiousdisorders, a dosage regimen of 4 times daily or less is preferred and adosage regimen of 1 or 2 times daily is particularly preferred.

It will be understood, however, that the specific dose level for anyparticular patient will depend upon a variety of factors including theactivity of the specific compound employed, the age, body weight,general health, sex, diet, time of administration, route ofadministration, and rate of excretion, drug combination and the severityof the particular disease undergoing therapy.

Combination Administration

The compounds of the invention may also be useful in combination withother pharmaceutically active agents such as antibacterial agents,antiviral agents, antifungal agents, anti-inflammatories, interferon,efflux-pump inhibitors, and beta-lactamase inhibitors. Antibiotic agentsinclude any molecule that tends to prevent, inhibit or destroy life andas such, includes anti-bacterial agents, anti-fungicides, anti-viralagents, and anti-parasitic agents.

A composition comprising a compound or salt of Formula I or Formula I incombination with another one or more antibacterial agent, antiprotozoalagent, antifungal agent, antiviral agent, interferon, efflux-pumpinhibitor, or beta-lactamase inhibitor is provided herein.

Pharmaceutical compositions of the invention include single dosage formscontaining of a compound of Formula I and/or Formula II and one or moreother active agent, dosage forms containing more than one compound ofFormula I and/or Formula II, and separate administration of a compoundof Formula I and/or Formula II with another active agent.

The following active agents, which are useful in combinations of theinvention, may be isolated from an organism that produces the agent orsynthesized by methods known to those of ordinary skill in the art ofmedicinal chemistry or purchased from a commercial source.

Anti-bacterial antibiotic agents include, but are not limited to,penicillins, cephalosporins, carbacephems, cephamycins, carbapenems,monobactams, aminoglycosides, glycopeptides, quinolones, tetracyclines,macrolides, and fluoroquinolones (see Table below). Examples ofantibiotic agents include, but are not limited to, Penicillin G (CASRegistry No.: 61-33-6); Methicillin (CAS Registry No.: 61-32-5);Nafcillin (CAS Registry No.: 147-52-4); Oxacillin (CAS Registry No.:66-79-5); Cloxacillin (CAS Registry No.: 61-72-3); Dicloxacillin (CASRegistry No.: 3116-76-5); Ampicillin (CAS Registry No.: 69-53-4);Amoxicillin (CAS Registry No.: 26787-78-0); Ticarcillin (CAS RegistryNo.: 34787-01-4); Carbenicillin (CAS Registry No.: 4697-36-3);Mezlocillin (CAS Registry No.: 51481-65-3); Azlocillin (CAS RegistryNo.: 37091-66-0); Piperacillin (CAS Registry No.: 61477-96-1); Imipenem(CAS Registry No.: 74431-23-5); Aztreonam (CAS Registry No.:78110-38-0); Cephalothin (CAS Registry No.: 153-61-7); Cefazolin (CASRegistry No.: 25953-19-9); Cefaclor (CAS Registry No.: 70356-03-5);Cefamandole formate sodium (CAS Registry No.: 42540-40-9); Cefoxitin(CAS Registry No.: 35607-66-0); Cefuroxime (CAS Registry No.:55268-75-2); Cefonicid (CAS Registry No.: 61270-58-4); Cefmetazole (CASRegistry No.: 56796-20-4); Cefotetan (CAS Registry No.: 69712-56-7);Cefprozil (CAS Registry No.: 92665-29-7); Loracarbef (CAS Registry No.:121961-22-6); Cefetamet (CAS Registry No.: 65052-63-3); Cefoperazone(CAS Registry No.: 62893-19-0); Cefotaxime (CAS Registry No.:63527-52-6); Ceftizoxime (CAS Registry No.: 68401-81-0); Ceftriaxone(CAS Registry No.: 73384-59-5); Ceftazidime (CAS Registry No.:72558-82-8); Cefepime (CAS Registry No.: 88040-23-7); Cefixime (CASRegistry No.: 79350-37-1); Cefpodoxime (CAS Registry No.: 80210-62-4);Cefsulodin (CAS Registry No.: 62587-73-9); Fleroxacin (CAS Registry No.:79660-72-3); Nalidixic acid (CAS Registry No.: 389-08-2); Norfloxacin(CAS Registry No.: 70458-96-7); Ciprofloxacin (CAS Registry No.:85721-33-1); Ofloxacin (CAS Registry No.: 82419-36-1); Enoxacin (CASRegistry No.: 74011-58-8); Lomefloxacin (CAS Registry No.: 98079-51-7);Cinoxacin (CAS Registry No.: 28657-80-9); Doxycycline (CAS Registry No.:564-25-0); Minocycline (CAS Registry No.: 10118-90-8); Tetracycline (CASRegistry No.: 60-54-8); Amikacin (CAS Registry No.: 37517-28-5);Gentamicin (CAS Registry No.: 1403-66-3); Kanamycin (CAS Registry No.:8063-07-8); Netilmicin (CAS Registry No.: 56391-56-1); Tobramycin (CASRegistry No.: 32986-56-4); Streptomycin (CAS Registry No.: 57-92-1);Azithromycin (CAS Registry No.: 83905-01-5); Clarithromycin (CASRegistry No.: 81103-11-9); Erythromycin (CAS Registry No.: 114-07-8);Erythromycin estolate (CAS Registry No.: 3521-62-8); Erythromycin ethylsuccinate (CAS Registry No.: 41342-53-4); Erythromycin glucoheptonate(CAS Registry No.: 23067-13-2); Erythromycin lactobionate (CAS RegistryNo.: 3847-29-8); Erythromycin stearate (CAS Registry No.: 643-22-1);Vancomycin (CAS Registry No.: 1404-90-6); Teicoplanin (CAS Registry No.:61036-64-4); Chloramphenicol (CAS Registry No.: 56-75-7); Clindamycin(CAS Registry No.: 18323-44-9); Trimethoprim (CAS Registry No.:738-70-5); Sulfamethoxazole (CAS Registry No.: 723-46-6); Nitrofurantoin(CAS Registry No.: 67-20-9); Rifampin (CAS Registry No.: 13292-46-1);Mupirocin (CAS Registry No.: 12650-69-0); Metronidazole (CAS RegistryNo.: 443-48-1); Cephalexin (CAS Registry No.: 15686-71-2); Roxithromycin(CAS Registry No.: 80214-83-1); Co-amoxiclavuanate; combinations ofPiperacillin and Tazobactam; and their various salts, acids, bases, andother derivatives.

Anti-fungals agents include but are not limited to Amphotericin B,Candicidin, Dermostatin, Filipin, Fungichromin, Hachimycin, Hamycin,Lucensomycin, Mepartricin, Natamycin, Nystatin, Pecilocin, Perimycin,Azaserine, Griseofulvin, Oligomycins, Neomycin, Pyrrolnitrin, Siccanin,Tubercidin, Viridin, Butenafine, Naftifine, Terbinafine, Bifonazole,Butoconazole, Chlordantoin, Chlormidazole, Cloconazole, Clotrimazole,Econazole, Enilconazole, Fenticonazole, Flutrimazole, Isoconazole,Ketoconazole, Lanoconazole, Miconazole, Omoconazole, Oxiconazole,Sertaconazole, Sulconazole, Tioconazole, Tolciclate, Tolindate,Tolnaftate, Fluconawle, Itraconazole, Saperconazole, Terconazole,Acrisorcin, Amorolfine, Biphenamine, Bromosalicylchloranilide,Buclosamide, Calcium Propionate, Chlorphenesin, Ciclopirox, Cloxyquin,Coparaffinate, Diamthazole, Exalamide, Flucytosine, Halethazole,Hexetidine, Loflucarban, Nifuratel, Potassium Iodide, Propionic Acid,Pyrithione, Salicylanilide, Sodium Propionate, Sulbentine,Tenonitrozole, Triacetin, Ujothion, Undecylenic Acid, and ZincPropionate.

Antiviral agents include, but are not limited to, Acyclovir, Cidofovir,Cytarabine, Dideoxyadenosine, Didanosine, Edoxudine, Famciclovir,Floxuridine, Ganciclovir, Idoxuridine, Inosine Pranobex, Lamivudine,MADU, Penciclovir, Sorivudine, Stavudine, Trifluridine, Valacyclovir,Vidarabine, ZaIcitabine, Zidovudine, Acemannan, Acetylleucine,Amantadine, Amidinomycin, Delavirdine, Foscarnet, Indinavir,Interferon-alpha, Interferon-beta, Interferon-gamma, Kethoxal, Lysozyme,Methisazone, Moroxydine, Nevirapine, Podophyllotoxin, Ribavirin,Rimantadine, Ritonavir2, Saquinavir, Stailimycin, Statolon,Tromantadine, and Xenazoic Acid.

Antiinflammatory agents include, but are not limited to, Enfenamic Acid,Etofenamate, FIufenamic Acid, Isonixin, Meclofenamic Acid, MefenamicAcid, Niflumic Acid, Talniflumate, Terofenamate, Tolfenamic Acid,Aceclofenac, Acemetacin, Alclofenac, Amfenac, Amtolmetin Guacil,Bromfenac, Bufexamac, Cinmetacin, Clopirac, Diclofenac, Etodolac,Felbinac, Fenclozic Acid, Fentiazac, Glucametacin, Ibufenac,Indomethacin, Isofezolac, Isoxepac, Lonazolac, Metiazinic Acid,Mofezolac, Oxametacine, Pirazolac, Proglumetacin, Sulindac, Tiaramide,Tolmetin, Tropesin, Zomepirac, Bumadizon, Butibufen, Fenbufen, Xenbucin,Clidanac, Ketorolac, Tinoridine, Alminoprofen, Benoxaprofen,Bermoprofen, Bucloxic Acid, Carprofen, Fenoprofen, Flunoxaprofen,Flurbiprofen, Ibuprofen, Ibuproxam, Indoprofen, Ketoprofen, Loxoprofen,Naproxen, Oxaprozin, Piketoprofen, Pirprofen, Pranoprofen, ProtizinicAcid, Suprofen, Tiaprofenic Acid, Ximoprofen, Zaltoprofen, Difenamizole,Epirizole, Apazone, Benzpiperylon, Feprazone, Mofebutazone, Morazone,Oxyphenbutazone, Phenylbutazone, Pipebuzone, Propyphenazone,Ramifenazone, Suxibuzone, Thiazolinobutazone, Acetaminosalol, Aspirin,Benorylate, Bromosaligenin, Calcium Acetylsalicylate, Diflunisal,Etersalate, Fendosal, Gentisic Acid, Glycol Salicylate, ImidazoleSalicylate, Lysine Acetylsalicylate, Mesalamine, Morpholine Salicylate,I-Naphthyl Salicylate, Olsalazine, Parsalmide, Phenyl Acetylsalicylate,Phenyl Salicylate, Salacetamide, Salicylamide O-Acetic Acid,Salicylsulfuric Acid, Salsalate, Sulfasalazine, Ampiroxicam, Droxicam,Isoxicam, Lornoxicam, Piroxicam, Tenoxicam, epsilon-AcetamidocaproicAcid, S-Adenosylmethionine, 3-Amino-4-hydroxybutyric Acid, Amixetrine,Bendazac, Benzydamine, alpha-Bisabolol, Bucolome, Difenpiramide,Ditazol, Emorfazone, Fepradinol, Guaiazulene, Nabumetone, Nimesulide,Oxaceprol, Paranyline, Perisoxal, Proquazone, Superoxide Dismutase,Tenidap, Zileuton, 21-Acetoxypregnenolone, Alclometasone, Algestone,Amcinonide, Beclomethasone, Betamethasone, Budesonide, Chloroprednisone,Clobetasol, Clobetasone, Clocortolone, Cloprednol, Corticosterone,Cortisone, Cortivazol, Deflazacort, Desonide, Desoximetasone,Dexamethasone, Diflorasone, Diflucortolone, Difluprednate, Enoxolone,Fluazacort, Flucloronide, Flumethasone, Flunisolide, FluocinoloneAcetonide, Fluocinonide, Fluocortin Butyl, Fluocortolone,Fluorometholone, Fluperolone Acetate, Fluprednidene Acetate,Fluprednisolone, Flurandrenolide, Fluticasone Propionate, Formocortal,Halcinonide, Halobetasol Propionate, Halometasone, Halopredone Acetale,Hydrocortamate, Hydrocortisone, Loteprednol Etabonale, Mazipredone,Medrysone, Meprednisone, Methylprednisolone, Mometasone Furoate,Paramethasone, Prednicarbate, Prednisolone, Prednisolone25-Diethylamino-acetate, Prednisolone Sodium Phosphate, Prednisone,Prednival, Prednylidene, Rimexolone, Tixocortol, Triamcinolone,Triamcinolone Acetonide, Triamcinolone Benetonide, and TriamcinoloneHexacetonide.

Compounds of the invention may be combined with one or more Betalactamase inhibitor when used in combination with a beta-lactam classantibiotic, such as penicillin or cephalosporins. Beta-lactamaseinhibitors include, but are not limited to Clavulanic acid, Sulbactam,Sultamacillin, and Tazobactam.

Compounds of the invention may also be combined with one or more effluxpump inhibitor, such as a quinazolinone efflux pump inhibitors,d-ornithine-d-homophenylalanine-3-aminoquinoline,Phe-Arg-b-naphthylamide, propafenone, a phenothiazine or thioxantheneefflux pump inhibitor, 1-aza-9-oxafluorenes,N-[4-[2-(3,4-dihydro-6,7-dimethoxy-2(1H)-isoquinolinyl)ethyl]phenyl]-9,10-dihydro-5-methoxy-9-oxo-4-Acridinecarboxamide,reserpine, Milbemycin, Cinchonine, Verapamil,L-phenylalanyl-N-2-naphthalenyl-L-Argininamide (and analogs),5′-methoxyhydnocarpin-D, methylxanthines, FK506, a cyclosporine effluxpump inhibitor, Nocardamine and other siderophores, Amiodarone,Cyclosporin A, Ro11-2933 (DMDP), Quinidine, and the optical isomers ofPropranolol, Quinine (SQl) and Quinidine, Quinine-10,11-epoxide,Quercetin, Amitriptyline, Taxuspine C derivatives, Emodin, MC-002434;Agosterol A; Pheophorbide; pyridoquinolines such as2,2′-[(2,8,10-trimethylpyrido[3,2-g]quinoline-4,6-diyl)bis(oxy)]bis[N,N-dimethyl-ethanamine,Gitonavir, and Gemfibrozil.

Synthesis of Compounds

The compounds of the invention are prepared according to methodswell-known to those skilled in the art of organic chemical synthesis.The starting materials used in preparing the compounds of the inventionare known, made by known methods, or are commercially available.

It is recognized that the skilled artisan in the art of organicchemistry can readily carry out standard manipulations of organiccompounds without further direction. Examples of such manipulations arediscussed in standard texts such as J. March, Advanced OrganicChemistry, John Wiley & Sons, 1992.

The skilled artisan will readily appreciate that certain reactions arebest carried out when other functionalities are masked or protected inthe compound, thus increasing the yield of the reaction and/or avoidingany undesirable side reactions. Often, the skilled artisan utilizesprotecting groups to accomplish such increased yields or to avoid theundesired reactions. These reactions are found in the literature and arealso well within the scope of the skilled artisan. Examples of many suchmanipulations can be found in, for example, T. Greene, Protecting Groupsin Organic Synthesis, John Wiley & Sons, 1981.

The compounds of the invention may have one or more chiral center. As aresult, one may selectively prepare one optical isomer, includingdiastereomers and enantiomers, over another, for example by chiralstarting materials, catalysts or solvents, or may prepare bothstereoisomers or both optical isomers, including diastereomers andenantiomers at once (a racemic mixture). Since the compounds of theinvention may exist as racemic mixtures, mixtures of optical isomers,including diastereomers and enantiomers, or stereoisomers may beseparated using known methods, such as through the use of, for example,chiral salts and chiral chromatography.

In addition, it is recognized that one optical isomer, including adiastereomer and enantiomer, or a stereoisomer, may have favorableproperties over the other. When a racemic mixture is discussed herein,it is clearly contemplated to include both optical isomers, includingdiastereomers and enantiomers, or one stereoisomer substantially free ofthe other.

The invention also includes also includes all energetically accessibleconformational and torsional isomers of the compounds disclosed.

When the substituent R7 in a compound of Formula I or Formula II isattached via an unsaturated aliphatic group, for example when R7 isphenyl (C2C6alkenyl), all geometric isomers of the compound areincluded.

This invention is further illustrated by the following examples thatshould not be construed as limiting. The contents of all references,patents and published patent applications cited throughout thisapplication are incorporated herein by reference.

EXAMPLES Abbreviations

The following abbreviations are used in the reaction schemes andsynthetic examples, which follow. This list in not meant to be anall-inclusive list of abbreviations used in the application asadditional standard abbreviations, which are readily understood by thoseskilled in the art of organic synthesis, may also be used in thesynthetic schemes and examples

(Boc)₂O—Di-t-butyl dicarbonaten-BuLi—n-Butyl lithium

DMAP—4-Dimethylaminopyridine DMF—N,N-DimethylformamideDMSO—Dimethylsulfoxide

EtOAc—Ethyl acetate

NBS—N-bromosuccinamide NCS—N-chlorosuccinamidePd(PPh₃)₄—Tetrakis(triphenylphosphine)palladium(0)

PTLC—Preparative thin layer chromatography

THF—Tetrahydrofuran

TLC—Thin-layer chromatography

General Methods

All nonaqueous reactions are performed under an atmosphere of dry argongas (99.99%) using oven- or flame-dried glassware. Microwave-assistedsyntheses are conducted in a commercial microwave reactor (DiscoverSystem, CEM Corporation). The progress of reactions is monitored usingthin-layer chromatography on glass plates coated with Merck silica gel60 (F₂₅₄). Flash column chromatography is performed on Merck silica gel60 (230-400 mesh). Melting points are recorded on an ElectrothermalModel IA9100 digital melting point apparatus; the reported values arethe average of three measurements. NMR spectra are recorded at ambienttemperature using a Bruker Avance 300 spectrometer (¹H at 300.1 MHz, ¹³Cat 75.5 MHz, and ¹⁹F at 282.4 MHz). The chemical shifts for ¹ H and ¹³Care reported in parts per million (δ) relative to externaltetramethylsilane and are referenced to signals of residual protons inthe deuterated solvent. The chemical shifts for ¹⁹F are reported inparts per million (δ) relative to external fluorotrichloromethane.Assignment of ¹H and ¹³C NMR data is based on extensive two-dimensionalcorrelation experiments (¹H-¹H COSY, ¹H-¹³C HMQC, ¹H-¹³C HMBC, and ¹H-¹HNOESY) and the usual principles of NMR spectroscopy (the magnitudes ofcoupling constants and chemical shifts). Analytical HPLC is performedusing a YMC Pack Pro C18 50×4.6 mm 5 μm column with an isocratic elutionof 0.24 min at 90:10 H₂O:CH₃CN containing 0.1% TFA followed by a 4-minlinear gradient elution from 90:10 to 10:90 at a flow rate of 2.5 mL/minwith UV detection at 254 nm. Preparative HPLC is performed using a YMCPack Pro C18 150×20.0 mm 5 μum column with an isocratic elution of 0.24min at 97:3 H₂O:CH₃CN containing 0.1% TFA followed by a 10-min lineargradient elution from 97:3 to 0:100 at a flow rate of 18.0 mL/min withUV detection at 254 nm. Low-resolution mass spectra are recorded on aThermo Finnigan Surveyor MSQ instrument (operating in APCI mode)equipped with a Gilson liquid chromatograph. Unless noted otherwise, thequasi-molecular ions, [M+H]⁺, observed in the low-resolution massspectra are the base peaks. High-resolution mass spectrometric analyses(ESI using sodium iodide as internal standard) are performed at the W.M. Keck Foundation Biotechnology Resource Laboratory (Yale University,New Haven, Conn.); the reported exact masses are the average of fivemeasurements. Elemental analysis is performed at Prevalere LifeSciences, Inc. (Whitesboro, N.Y.).

Example 1 General Method for Tthe Preparation of9-cyclopropyl-6-fluoro-7-phenyl-9H-isothiazolo[5,4-B]quinoline-3,4-diones(9)

9-cyclopropyl-6-fluoro-7-phenyl-9H-isothiazolo[5,4-b]quinoline-3,4-diones(9) are prepared in accordance with the synthetic scheme set forthbelow.

STEP A. Preparation of 4-bromo-2,5-difluorobenzoic acid (2)

Freshly titrated n-butyl lithium (27.0 ml, 1.39 M in hexanes) is addedslowly (over about 30 minutes) to a −78° C. solution of diethyl ether(90 ml) containing 1,4-dibromo-2,5-difluorobenzene (1, 10.22 g, 0.038mol). The resulting yellow solution is stirred at −78° C. for 2 hours togive a yellow suspension. Several pellets (˜10) of dry ice are added tothe suspension, which is then allowed to warm slowly to room temperatureas it degasses (approximately 40 minutes). The resulting suspension isacidified with a 1 M aqueous solution of hydrochloric acid (500 ml), andthe product extracted with diethyl ether (5×200 ml). The combinedorganics are washed with water (4×100 ml) and filtered. The ethersolution is concentrated to approximately 200 ml under reduced pressure,and the product extracted into a saturated aqueous solution of sodiumbicarbonate (3×200 ml). The combined aqueous extracts are washed withmethylene chloride (3×100 ml) and acidified with hydrochloric acid. Theproduct is extracted with diethyl ether (3×200 ml), and the combinedorganic extracts washed with water (2×200 ml), dried over magnesiumsulfate, and concentrated under reduced pressure to give (2) as a paleyellow solid. ¹H NMR (300 MHz, DMSO-d₆): δ7.74 (dd, J_(H-F)=8.5 Hz, 6.5Hz, 1H), 7.84 (dd, J_(H-F)=10.0 Hz, 5.5 Hz, 1H), 13.7 (br, 1H, CO₂H).¹⁹F{¹H} NMR (282 MHz, DMSO-d₆): δ-114.0 (d, J_(F-F)=17.0 Hz, 1F), −113.6(d, J_(F-F)=17.0 Hz, 1F). ¹³C{¹H} NMR (75 MHz, DMSO-d₆): δ113.6 (dd,J_(C-F)=23.5 Hz, 10.0 Hz), 118.4 (dd, J_(C-F)=26.5 Hz, 2.5 Hz, CH),120.0 (dd, J_(C-F)=19.0 Hz, 12.0 Hz), 122.2 (d, J_(C-F)=28.0 Hz, CH),154.4 (dd, J_(C- F)=245.0 Hz, 5.5 Hz, CF), 156.8 (dd, J_(C-F)=251.5 Hz,4.0 Hz, CF), 163.4 (m, CO₂H).

STEP B. Preparation of 3-(4-bromo-2,5-difluoro-phenyl)-3-oxo-propionicacid ethyl ester (3)

4-Bromo-2,5-difluorobenzoyl chloride is prepared from 2 as describedpreviously. [Reuman, M.; et. al, J. Med. Chem. (1995) 38, 2531-2540].Note that the addition of dimethylformamide is omitted from thisprocedure. This intermediate is used to prepare 3 as describedpreviously [Wierenga, W.; Skulnick, H. I. J. Org. Chem. 1979, 44,310-311]. ¹H NMR (300 MHz, CDCl₃): (enol, major) δ1.32 (t, J_(H-H)=7.0Hz, 3H, CO₂CH₂CH₃), 4.26 (q, J_(H-H)=7.0 Hz, 2H, CO₂CH₂CH₃), 5.85 (s,1H, CH₃C(OH)═CH—CO₂CH₂CH₃), 7.34 (dd, J_(H-F)=10.5 Hz, 5.5 Hz, 1H,aromatic), 7.64 (dd, J_(H-F)=9.0 Hz, 6.5 Hz, 1H, aromatic), 12.65 (s,1H, OH). ¹⁹F{¹H} NMR (282 MHz, CDCl₃): δ−114.8 (d, J_(F-F)=17.0 Hz, 1F),−112.6 (d, J_(F-F)=17.0 Hz, 1F). 3 (keto, minor): ¹H NMR (300 MHz,CDCl₃): δ1.24 (t, J_(H-H)=7.0 Hz, 3H, CO₂CH₂CH₃), 3.93 (d, J_(H-F)=4.0Hz, 2H, CH₂CO₂CH₂CH₃), 4.19 (q, J_(H-H)=7.0 Hz, 2H, CO₂CH₂CH₃), 7.40(dd, J_(H-F)=9.5 Hz, 5.5 Hz, 1H, aromatic), 7.68 (dd, J_(H-F)=8.5 Hz,6.0 Hz, 1H, aromatic). ¹⁹F{¹H} NMR (282 MHz, CDCl₃): δ−114.3 (d,J_(F-F)=17.0 Hz, 1F), −111.7 (d, J_(F-F)=17.0 Hz, 1F).

STEP C. Preparation of2-(4-bromo-2,5-difluoro-benzoyl)-3-cyclopropylamino-3-methylsulfanyl-acrylicacid ethyl ester (4)

Cyclopropyl thioisocyanate (0.57 ml, 6.15 mmole, 1.7 equiv.) is added toa stirred solution of 3-(4-Bromo-2,5-difluoro-phenyl)-3-oxo-propionicacid ethyl ester (3, 1.06 g, 3.5 mmole) in DMF (anhydrous, 10 ml) underargon at room temperature. The reaction mixture is cooled in an ice bathand NaH (150 mg, 60% in mineral oil, 3.7 mmole, 1.07 equiv.) is addedportionwise at 0-5° C. under argon. After addition, the reaction mixtureis allowed to warm to room temperature and stirred at room temperatureuntil TLC indicates there is no remaining starting material. CH₃I (0.38ml, 5.6 mmole, 1.7 equiv.) is then added to the reaction mixture. Thereaction is diluted with EtOAc and quenched with NH₄Cl solution afterstirring at room temperature for about 4 hours (TLC and LC MS are usedto determine reaction completion). The organics are washed with brine,dried over Na₂SO₄, and concentrated. The resulting crude oil (4, 1.6 g)is purified by column chromatography (Silica gel, 40% EtOAc in hexanes,gradient, 40 minutes) to yield 4 as a yellow oil. ¹H NMR (CDCl₃) δ: 7.15(m, 2H), 3.89 (q, 2H), 2.96 (m, 1H), 2.48 (s, 3H), 0.85 (m, 7H).

STEP D. Preparation of7-bromo-1-cyclopropyl-6-fluoro-2-methylfulfanyl-4-oxo-1,4-dihydro-quinoline-3-carboxylic acid ethyl ester (5)

NaH (345 mg, 60% in mineral oil, 8.6 mmole, 1.05 equiv.) is added to astirred solution of2-(4-bromo-2,5-difluoro-benzyl)-3-cyclopropylamino-3-methylsulfanyl-acrylicacid ethyl ester (4, 3.46 g, 8.2 mmole) in DMF (anhydrous, 100 ml). Thereaction mixture is stirred at 75° C. for 18 hours (TLC is used toindicate reaction completion). The reaction mixture is cooled, dilutedwith NH₄Cl solution, and extracted with EtOAc. The organics are washedwith brine (4×30 ml), dried over Na₂SO₄, and concentrated in vacuo togive 5 as a light yellow solid. This intermediate is used withoutfurther purification ¹H NMR (CDCl₃) indicated >98% purity). ¹HNMR(CDCl₃) δ: 8.09 (d, 1H), 7.90 (d, 1H), 4.35 (q, 2H), 3.22 (m, 1H), 2.49(s, 3H), 1.3-1.5 (m, 7H).

STEP E. Preparation of7-bromo-1-cyclopropyl-6-fluoro-2-mercapto-4-oxo-1,4-dihydroquinoline-3-carboxylicacid ethyl ester. (6)

Sodium hydrogen sulfide (5 mg, 0.09 mmole, 1.5 equiv.) is added to astirred solution of7-Bromo-1-cyclopropyl-6-fluoro-2-methylsulfanyl-4-oxo-1,4-dihydroquinoline-3-carboxylicacid ethyl ester (5, 24 mg, 0.06 mmole) in THF (tetrahydrofuran, 2 ml)under argon at room temperature. The reaction is then stirred at 45° C.until TLC indicated completion. The reaction mixture is diluted withwater and washed with diethyl ether. The aqueous layer is acidified by1N HCl to a pH of approximately 2, and extracted with EtOAc. Theresulting organics are washed with brine, dried over Na₂SO₄, andconcentrated in vacuo. The crude product is purified by PTLC (20% CH₃OHin CHCl₃) to give 6. Alternatively, sodium hydrogen sulfide (20 mg, 0.36mmole, 1.5 equiv.) is added to a stirred solution of 5 (crude, 96 mg,0.24 mmole) in DMF (6 ml) under argon at room temperature. The reactionis then stirred at 40° C. until TLC indicates completion. The reactionmixture is diluted with water, acidified by 1N HCl to a pH ofapproximately 2, and extracted with EtOAc. The resulting organics arewashed with brine, dried over Na₂SO₄, and concentrated. The crude ispurified by PTLC (20% CH₃OH in CHCl₃) to yield 6. ¹H NMR (CDCl₃) δ: 8.40(d, 1H), 8.06 (d, 1H), 4.71 (q, 2H), 3.46 (m, 1H), 1.68 (m, 7H).

STEP F. Preparation of7-bromo-9-cyclopropyl-6-fluoro-9H-isothiazolo[5,4-B]quinoline-3,4-dione(7)

A mixture of NaHCO₃ solution (51 mg, 0.9 ml water) andhydroxylamine-O-sulfonic acid (27 mg, 0.24 mmole, 4.2 equiv.) is addedto a stirred solution of7-Bromo-1-cyclopropyl-6-fluoro-2-mercapto-4-oxo-1,4-dihydroquinoline-3-carboxylicacid ethyl ester (6, 22 mg, 0.057 mmole) in THF (0.7 ml). The reactionmixture is stirred at room temperature for approximately 3 hours untilthe reaction is complete. The reaction mixture is acidified by theaddition of 0.5N HCl and filtered. The resulting solid is washed withwater (3×) and dried yielding 7 as a white solid. ¹H NMR (DMSO-d₆) δ:8.39 (d, 1H), 8.06 (d, 1H), 3.63 (m, 1H), 1.41 (m, 2H), 1.26 (m, 2H).¹⁹F NMR (DMSO-d₆) δ: 114.9 (s, 1H).

STEP G. Preparation of tert-butyl7-bromo-9-cyclopropyl-6-fluoro-3,4-dioxoisothiazolo[5,4-B]quinoline-2(3H,4H,9H)-carboxylate(8)

4-Dimethylaminopyridine (DMAP, catalytic amount) and (Boc)₂O (27 mg, 2equivalents, 0.12 mmole) are added to a stirred solution of7-Bromo-9-cyclopropyl-6-fluoro-9H-isothiazolo[5,4-b]quinoline-3,4-dione(7, 22 mg, 0.062 mmole) in DMF (0.75 ml) under nitrogen. The reaction isstirred at room temperature for 18 hours. Water (1 ml) is added to thereaction mixture and the solid filtered, washed with water, and dried toyield 8 as a white solid. ¹H NMR (DMSO-d₆) δ: 8.30 (d, 1H), 7.91 (d,1H), 3.58 (m, 1H), 1.57 (s, 9H), 1.41 (m, 2H), 1.27 (m, 2H). ¹⁹F NMR(DMSO-d₆)) δ: 111.1 (s, 1H).

STEP H. Preparation of9-cyclopropyl-6-fluoro-7-aryl-9H-isothiazolo[5,4-B]quinoline-3,4-dione(9)

Pd(PPh₃)₄ (6 to 10 mole %) is added to a stirred suspension of7-Bromo-9-cyclopropyl-6-fluoro-3,4-dioxo-4,9-dihydro-3H-isothiazolo[5,4-b]quindine-2-carboxylicacid tert-butyl ester (8, 20 mg, 0.044 mmole) in DMF (1 ml), followed byaddition of a boronic acid (2 equivalents, 0.088 mmole) and NaHCO₃solution (1M, 0.2 ml, 4.5 equiv.) under argon at room temperature. Thereaction tube is sealed and then stirred in a microwave (100W, 130° C.)until completion (usually 10 minutes, though a longer reaction time maybe needed for some boronic acids). The reaction mixture is monitored byLC MS until no starting material remains. The reaction mixture is thenfiltered and the filtrate concentrated in vacuo. The residue is washedwith a mixture of MeOH: diethyl ether (approximately 5:95, 3×). Theresulting light yellow solid product is dried and analyzed. Someproducts require HPLC purification.

Alternate Procedure for the Synthesis of9-cyclopropyl-6-fluoro-7-aryl-9H-isothiazolo[5,4-B]quinoline-3,4-dione

Pd (PPh₃)₄ (3.1 mg, 6% mole), boronic acid or ester (2 equiv., 0.088mmole) and NaHCO₃ solution (1M, 0.2 mL, 4.5 equiv.) under argon at roomtemperature is added to a stirred solution of7-bromo-9-cyclopropyl-6-fluoro-9H-isothiazolo[5,4-b]quinoline-3,4-dione(20 mg, 0.044mmole) in DMF (1 mL). The reaction mixture is degassed bybubbling argon through for 10 minutes at room temperature. The reactiontube is sealed and then heated in a microwave (100W, 130° C.) until thereaction reaches completion (10-20 minutes). The reaction mixture iscooled to room temperature and then filtered. The filtrate isconcentrated in vacuo. The residue is dissolved in a mixture ofDMF:CHCl₃:MeOH (0.5:3:0.5) (4 mL) and precipitated with diethyl ether.This dissolution and precipitation step is repeated 5 times. Theresulting light yellow solid is washed with water (3 mL) and dried toafford the title compound.

Example 2 General Method for the Preparation of9-cyclopropyl-8-methoxy-7-aryl-9H-isothiazolo[5,4-B]quinoline-3,4-diones

9-Cyclopropyl-8-methoxy-7-aryl-9H-isothiazolo[5,4-b]quinoline-3,4-diones(18) are prepared in accordance with the synthetic scheme set forthbelow.

STEP A. Preparation of 3-fluoro-2-methoxyphenylamine (11)

Potassium carbonate (59.25 g, 0.43 mol) is added slowly to a solution ofdimethylformamide (200 mL) containing 2-fluoro-6-nitrophenol (10, 33.63g, 0.21 mol) and dimethylsulfate (41.0 mL, 0.43 mol) at roomtemperature. The orange mixture is stirred at 80° C. for 6 h. Theresulting yellow mixture is cooled to room temperature, diluted withwater (500 mL), and extracted with hexanes (3×500 mL). The combinedorganic extracts are dried over magnesium sulfate and evaporated underreduced pressure to give 1-fluoro-2-methoxy-3-nitrobenzene as a yellowoil. This product is of sufficient purity (≧95% by NMR spectroscopy) touse directly in the next synthetic step. ¹H NMR (CDCl₃): δ4.08 (d,J_(H-F)=2.0 Hz, 3H, OCH₃), 7.13 (apparent t of d, J_(H-H)=8.5 Hz,J_(H-F)=5.0 Hz, 1H, H-5), 7.34 (d, J_(H-F)=10.5 Hz, J_(H-H)=8.5 Hz,J_(H-H)=1.5 Hz, 1H, H-6), 7.58 (d of apparent t, J_(H-H)=8.5 Hz,J_(H-H)=1.5 Hz, J_(H-F)=1.5 Hz, 1H, H-4). ¹⁹F{¹H} NMR (CDCl₃): δ−126.7(s). ¹³C{¹H} NMR (CDCl₃): δ62.6 (d, J_(C-F)=5.5 Hz, OCH₃), 120.2 (d,J_(C-F)=3.5 Hz, C-4), 121.1 (d, J_(C-F)=19.5 Hz, C-6), 123.2 (d,J_(C-F)=8.0 Hz, C-5), 142.2 (d, J_(C-F)=14.5 Hz, C-2), 144.8 (br, C-3),156.2 (d, J_(C-F)=251.5 Hz, C-1). LCMS m/z calcd for C₇H₆FNO₃ ([M]⁺)171; found 183 ([M−CH₂O+H+CH₃CN]⁺, 26%), 183 ([M−CH₂O +H]⁺, 100%).

A mixture containing 1-fluoro-2-methoxy-3-nitrobenzene (36.30 g, 0.21mol), palladium on carbon (10% w/w, ˜8 g), and methanol (200 mL) isstirred under an atmosphere of hydrogen (1 atm) for 27 h. The mixture isfiltered and the resulting solution is evaporated to dryness underreduced pressure to give 11 as a brown oil. This product is ofsufficient purity (≧95% by NMR spectroscopy) to use directly in the nextsynthetic step. ¹H NMR (CDCI₃): δ3.75 (br, 2H, NH₂), 3.91 (d,J_(H-F)=1.5 Hz, 3H, OCH₃), 6.46 (m, 2H, overlapping H-4 and H-6), 6.79(apparent t of d, J_(H-H)=8.0 Hz, J_(H-F)=5.5 Hz, 1H, H-5). ¹⁹F{¹H} NMR(CDCl₃): δ−132.5 (s). ¹³C{¹H} NMR (CDCl₃): δ60.7 (d, J_(C-F)=5.0 Hz,OCH₃), 106.1 (d, J_(C-F)=19.5 Hz, C-4), 110.9 (d, J_(C-F)=2.5 Hz, C-6),123.7 (d, J_(C-F)=9.5 Hz, C-5), 134.9 (d, J_(C-F)=13.0 Hz, C-2), 141.3(d, J_(C-F)=5.0 Hz, C-1), 154.4 (d, J_(C-F)=244.0 Hz, C-3). LCMS m/zcalcd for C₇H₈FNO ([M]⁺) 141; found 142 ([M+H]⁺).

STEP B. Preparation of 4-bromo-2-fluoro-3-methoxybenzoic acid (12).

(a) Hydrobromic acid (48% in water, 140 mL) is added slowly to analiquot of 11 (14.33 g, 101.5 mmol) cooled to 0° C. The resulting solidis broken up with a glass rod and stirred vigorously at 0° C. for 10min. A solution of sodium nitrite (7.40 g, 107.2 mmol) in water (50 mL)is added slowly (˜1.5 h) to the stirred slurry containing3-fluoro-2-methoxyphenylamine and hydrobromic acid, maintaining thetemperature of the reaction mixture below 5° C. A purple solution ofcuprous bromide (9.62 g, 67.1 mmol) in hydrobromic acid (48% in water,50 mL) is added dropwise to the reaction mixture, maintaining thetemperature of the reaction mixture below 5° C. The resulting reactionmixture is heated at 60° C. until the evolution of gas ceases (˜2.5 h).The reaction mixture is cooled to room temperature, and the productextracted with diethyl ether (6×150 mL). The combined organic extractsare washed with brine (3×150 mL), dried over magnesium sulfate, andevaporated under reduced pressure to give1-bromo-3-fluoro-2-methoxybenzene as a brown oil. This product is ofsufficient purity (≧95% by NMR spectroscopy) to use directly in the nextsynthetic step. ¹H NMR (CDCl₃): δ3.95 (d, J_(H-F)=1.5 Hz, 3H, OCH₃),6.88 (apparent t of d, J_(H-H)=8.0 Hz, J_(H-F)=5.5 Hz, 1H, H-5), 7.04(d, J_(H-F)=10.5 Hz, J_(H-H)=8.0 Hz, J_(H-H)=1.5 Hz, 1H, H-4), 7.30 (dof apparent t, J_(H-H)=8.0 Hz, J_(H-H)=1.5 Hz, J_(H-F)=1.5 Hz, 1H, H-6).¹⁹F{¹H} NMR (CDCl₃): δ−127.7 (s). ¹³C{¹H} NMR (CDCl₃): 61.4 (d,J_(C-F)=5.0 Hz, OCH₃), 116.2 (d, J_(C-F)=19.5 Hz, C-4), 117.7 (d,J_(C-F)=3.0 Hz, C-1), 124.5 (d, J_(C-F)=8.0 Hz, C-5), 128.5 (d,J_(C-F)=3.5 Hz, C-6), 145.7 (d, J_(C-F)=12.5 Hz, C-2), 156.2 (d,J_(C-F)=250.5 Hz, C-3).

(b) Lithium diisopropylamide (LDA) is formed by dropwise addition ofn-butyllithium (1.6 M in hexanes, 56.0 mL, 89.6 mmol) to a stirredsolution of diisopropylamine (13.7 mL, 96.9 mmol) in tetrahydrofuran(150 mL) at −78° C. The resulting solution is stirred at −78° C. for 5min, 0° C. for 15 min, and then cooled again to −78° C. A solution of1-bromo-3-fluoro-2-methoxybenzene (15.28 g, 74.5 mmol) intetrahydrofuran (40 mL) is added dropwise to the previous solution overa period of 30 min. to give an amber solution. After stirring thissolution at −78° C. for 1.5 h, dry ice (˜125 g) is added, and theresulting mixture is allowed to warm slowly (˜1 h) to room temperaturewith stirring as it degasses. The reaction mixture is acidified to pH ˜1by addition of a 5% aqueous solution of hydrochloric acid (˜500 mL), andthe product is extracted with diethyl ether (6×100 mL). The combinedorganic extracts are washed with brine (100 mL), and the productextracted with a saturated solution of aqueous sodium bicarbonate (3×100mL). The combined aqueous extracts (pH ˜9) are washed with diethyl ether(3×100 mL) and acidified slowly to pH ˜1 by addition of a 37% aqueoussolution of hydrochloric acid (˜50 mL). The product is extracted withdiethyl ether (3×200 mL), and the combined organic extracts are washedwith brine (100 mL), dried over anhydrous magnesium sulfate, andconcentrated under reduced pressure to give 12 as an off-white solid.This product is of sufficient purity (≧95% by NMR spectroscopy) to usedirectly in the next synthetic step. mp 168-170° C. ¹H NMR (CD₃OD):δ3.92 (d, J_(H-F)=1.0 Hz, 3H, OCH₃), 7.44 (dd, J_(H-H)=8.5 Hz,J_(H-F)=1.5 Hz, 1H, H-5), 7.55 (dd, J_(H-H)=8.5 Hz, J_(H-F)=7.0 Hz, 1H,H-6). ¹⁹F {¹H} NMR (CD₃OD): δ−127.0 (s). ¹³C{¹H} NMR (CD₃OD): δ62.1 (d,J_(C-F)=4.5 Hz, OCH₃), 121.5 (d, J_(C-F)=8.5 Hz, C-1), 123.6 (d,J_(C-F)=2.0 Hz, C-4), 128.0 (s, C-6), 129.0 (d, J_(C-F)=4.5 Hz, C-5),147.7 (d, J_(C-F)=13.5 Hz, C-3), 157.1 (d, J_(C-F)=263.5 Hz, C-2), 166.3(d, J_(C-F)=3.0 Hz, CO₂H). HRMS m/z calcd for C₈H₆ ⁷⁹BrFNaO₃ 270.9382([M+Na]⁺); found 270.9377.

STEP C. Preparation of ethyl3-(4-bromo-2-fluoro-3-methoxyphenyl)-3-oxopropionate (13).

Compound 13 is prepared using the general two-step method of Wierengaand Skulnick. (Wierenga, W.; Skulnick, H. I. J. Org. Chem. 1979, 44,310-311.)

(a) Dimethylformamide (5 drops) is added by Pasteur pipette to a mixturecontaining 12 (5.30 g, 21.3 mmol) and oxalyl chloride in methylenechloride (2.0 M, 21.3 mL, 42.6 mmol) at room temperature. The resultingmixture is stirred until an amber solution forms and the evolution ofgas ceases (1 h). The solution is concentrated under reduced pressure togive the intermediate acid chloride as an off-white solid that is useddirectly in the following step.

(b) n-Butyllithium (1.6 M in hexanes) is added to a cooled (−78° C.)solution of tetrahydrofuran (50 mL) containing ethyl hydrogen malonate(5.62, 42.5 mmol) and 2,2′-bipyridyl (8.2 mg as indicator). Thetemperature of the reaction mixture is allowed to rise to ˜0° C. duringthe addition of n-butyllithium. Sufficient n-butyllithium (˜50 mL) isadded until a pink color persists at −5° C. for 5-10 min. A solution ofthe acid chloride (vide supra) in methylene chloride (20 mL) is added inone portion to the reaction mixture that has been cooled to −78° C. Theresulting mixture is allowed to warm to 10° C. (˜30 min), and quenchedwith an aqueous solution of hydrochloric acid (1 M, 100 mL). Thereaction mixture is extracted with diethyl ether (3×100 mL). Thecombined organic extracts are washed with a saturated aqueous solutionof sodium bicarbonate (3×100 mL), followed by brine (100 mL), dried overmagnesium sulfate, and evaporated under reduced pressure to give thecrude product. This material is purified by flash column chromatography(eluting with ethyl acetate/hexanes (1:6 v/v); R_(f) 0.43) to give pure13 as a pale orange oil that solidified upon standing. mp 52-53° C. Thetitle compound exists as a mixture of keto (major) and enol (minor)tautomers at room temperature in CDCl₃, DMSO-d₆, and CD₃OD. ¹H NMR(CDCl₃): δ1.27 (t, J_(H-H)=7.0 Hz, keto CO₂CH₂CH₃), 1.34 (t, J_(H-H)=7.0Hz, enol CO₂CH₂CH₃), 3.96 (m, overlapping keto OCH₃, enol OCH₃, and ketoC(O)CH₂CO₂CH₂CH₃), 4.22 (q, J_(H-H)=7.0 Hz, keto CO₂CH₂CH₃), 4.27 (q,J_(H-H)=7.0 Hz, enol CO₂CH₂CH₃), 5.81 (d, J_(H-F)=0.5 Hz, enolC(OH)═CHCO₂CH₂CH₃), 7.39 (dd, J_(H-H)=8.5 Hz, J_(H-F)=1.5 Hz, enolaromatic H-5), 7.43 (dd, J_(H-H)=8.5 Hz, J_(H-F)=1.5 Hz, keto aromaticH-5), 7.47 (dd, J_(H-H)=8.5 Hz, J_(H-F)=7.0 Hz, enol aromatic H-6), 7.53(dd, J_(H-H)=8.5 Hz, J_(H-F)=7.0 Hz, keto aromatic H-6), 12.67 (s, enolOH). ¹⁹F{¹H} NMR (CDCl₃): δ−126.3 (s, enol), −125.9 (s, keto). LCMS m/zcalcd for C₁₂H₁₂ ⁷⁹BrFO₄ ([M]⁺) 318; found 319 ([M+H]⁺). HRMS m/z calcdfor C₁₂H₁₂ ⁷⁹BrFNaO₄ 340.9801 ([M+Na]⁺); found 340.9797.

STEP D. Preparation of ethyl3-(4-bromo-2-fluoro-3-methoxyphenyl)-2-(cyclopropyliminomethylsulfanylmethyl)-3-hydroxyacrylate(14).

Sodium hydride (60% in mineral oil, 73.7 mg, 1.92 mmol) is addedportionwise to a cooled (0° C.) solution containing 13 (569 mg, 1.78mmol), cyclopropyl isothiocyanate (500 μL, 5.40 mmol), anddimethylformamide (5.0 mL). The resulting mixture was allowed to warm toroom temperature with stirring overnight (18.5 h). Methyl iodide (700μL, 11.22 mmol) is added to the resulting solution to give a precipitatewithin minutes. The mixture is stirred for an additional 24 h. Thereaction mixture is quenched by addition of a saturated aqueous solutionof ammonium chloride (50 mL) and extracted with ethyl acetate (3×100mL). The combined organic extracts are washed with brine (200 mL), driedover magnesium sulfate, and evaporated under reduced pressure to givethe crude product. This material is purified by flash columnchromatography (eluting with 10% v/v ethyl acetate in methylenechloride; R_(f)0.59) to give 586.0 mg (76% yield) of 14 as a viscousyellow oil. ¹H NMR (CDCl₃): δ0.86 (m, 2H, c-Pr CH₂), 0.89 (t,J_(H-H)=7.0 Hz, CO₂CH₂CH₃), 0.98 (m, 2H, c-Pr CH₂), 2.52 (s, 3H, S—CH₃),3.01 (m, 1H, c-Pr CH), 3.90 (q, J_(H-H)=7.0 Hz, 2H, CO₂CH₂CH₃), 3.94 (d,J_(H-F)=1.5 Hz, 3H, OCH₃), 6.97 (dd, J_(H-H)=8.5 Hz, J_(H-F)=6.5 Hz, 1H,aromatic H-6), 7.30 (dd, J_(H-H)=8.5 Hz, J_(H-F)=1.5 Hz, 1H, aromaticH-5), 11.91 (br, 1H, OH). ¹⁹F{¹H} NMR (CDCl₃): δ−130.4 (s). ¹³C{¹H} NMR(CDCl₃): δ8.6 (c-Pr CH₂), 13.5 (CO₂CH₂CH₃), 18.1 (S—CH₃), 28.5 (c-PrCH), 60.3 (CO₂CH₂CH₃), 61.4 (d, J_(C-F)=5.0 Hz, OCH₃), 104.2(—C(OH)═C(CO₂CH₂CH₃)—), 118.3 (d, J_(C-F)=2.5 Hz, aromatic C-4), 123.4(d, J_(C-F)=3.5 Hz, aromatic C-6), 127.6 (d, J_(C-F)=3.5 Hz, aromaticC-5), 131.9 (d, J_(C-F)=14.5 Hz, aromatic C-1), 145.1 (d, J_(C-F)=13.5Hz, aromatic C-3), 152.6 (d, J_(C-F)=253.0 Hz, C-2), 167.7 (CO₂CH₂CH₃),174.5 (—N═C(S—CH₃)—), 185.5 (—C(OH)═C(CO₂CH₂CH₃)—). LCMS m/z calcd forC₁₇H₁₉ ⁷⁹BrFNO₄S ([M]⁺) 431; found 432 ([M +H]⁺). HRMS m/z calcd forC₁₇H₂₀ ⁷⁹BrFNO₄S 432.0280 ([M+H]⁺); found 432.0276.

STEP E. Preparation of ethyl7-bromo-1-cyclopropyl-8-methoxy-2-methylfulfanyl-4-oxo-1,4-dihydroquinoline-3-carboxylate(15).

Sodium hydride (60% in mineral oil, 51.9 mg, 1.30 mmol) is addedportionwise to a solution of 14 (527.6 mg, 1.22 mmol) indimethylformamide (5.0 mL) at room temperature. The reaction mixture isheated at 75° C. for 75 h, cooled to room temperature, and quenched byaddition of a saturated aqueous solution of ammonium chloride (75 mL).The mixture is extracted with ethyl acetate (3×75 mL). The combinedorganic extracts are washed with brine (75 mL), dried over magnesiumsulfate, and evaporated under reduced pressure to give crude 15 as a tansolid. This product is of sufficient purity (≧95% by NMR spectroscopy)to use directly in the next synthetic step. ¹H NMR (CDCl₃): δ0.70 (m,2H, c-Pr CH₂), 1.18 (m, 2H, c-Pr CH₂), 1.39 (t, J=7.0 Hz, 3H,CO₂CH₂CH₃), 2.63 (s, 3H, S—CH₃), 3.68 (m, 1H, c-Pr CH), 3.80 (s, 3H,OCH₃), 4.40 (q, J=7.0 Hz, 2H, CO₂CH₂CH₃), 7.54 (d, J=8.5 Hz, 1H,aromatic H-6), 7.88 (d, J=8.5 Hz, 1H, aromatic H-5). ¹³C{¹H} NMR(CDCl₃): δ12.4 (br, c-Pr CH₂), 14.2 (CO₂CH₂CH₃), 18.4 (S—CH₃), 37.0(c-Pr CH), 60.8 (OCH₃), 61.8 (CO₂CH₂CH₃), 122.7 (CH, C-5), 123.1 (C—Br,C-7), 123.6 (C-3), 129.2 (C-4a), 129.3 (CH, C-6), 140.0 (C-8a), 147.9(C—OCH₃, C-8), 156.3 (C—S—CH₃, C-2), 165.5 (CO₂CH₂CH₃), 173.6 (C═O,C-4). LCMS m/z calcd for C₁₇H₁₈ ⁷⁹BrNO₄S ([M]⁺) 411; found 412 ([M+H]⁺).HRMS m/z calcd for C₁₇H₁₈ ⁷⁹BrNNaO₄S 434.0038 ([M+Na]⁺); found 434.0031.

STEP F. ethyl7-bromo-1-cyclopropyl-2-methanesulfinyl-8-methoxy-4-oxo-1,4-dihydroquinoline-3-carboxylate(16)

m-Chloroperoxybenzoic acid (≦77%, 273.5 mg, 1.22 mmol) is added in oneportion to a solution of crude ethyl 15 (from above ˜1.22 mmol) inmethylene chloride (5.0 mL) at room temperature. The reaction mixture isstirred for 1 h, diluted with methylene chloride (10 mL), and washedwith a saturated aqueous solution of sodium bicarbonate (25 mL). Theorganic layer is dried over magnesium sulfate and evaporated underreduced pressure to give the crude product. This material is purified byflash column chromatography (eluting with ethyl acetate; R_(f)0.37) togive 290.9 mg of pure ethyl 16 as a white solid. ¹H NMR (CDCl₃): δ0.54(m, 1H, c-Pr CH₂ (A)), 0.93 (m, 1H, c-Pr CH₂ (B)), 1.12 (m, 1H, c-Pr CH₂(A)), 1.28 (m, 1H, c-Pr CH₂ (B)), 1.38 (t, J=7.0 Hz, 3H, CO₂CH₂CH₃),3.26 (s, 3H, S(O)—CH₃), 3.83 (s, 3H, OCH₃), 3.92 (m, 1H, c-Pr CH), 4.40(m, 2H, overlapping CO₂CHHCH₃), 7.58 (d, J=8.5 Hz, 1H, aromatic H-6),7.87 (d, J=8.5 Hz, 1H, aromatic H-5). ¹³C{¹H} NMR (CDCl₃): δ10.8 (br,c-Pr CH₂ (A)), 13.9 (br, c-Pr CH₂ (B)), 14.1 (CO₂CH₂CH₃), 35.1 (c-PrCH), 41.4 (S(O)—CH₃), 61.1 (OCH₃), 62.1 (CO₂CH₂CH₃), 118.9 (C-3), 122.8(CH, C-5), 123.9 (C—Br, C-7), 129.5 (C-4a), 130.0 (CH, C-6), 138.2(C-8a), 148.3 (C—OCH₃, C-8), 164.0 (CO₂CH₂CH₃), 164.1 (br, C-S(O)—CH₃,C-2), 174.6 (C═O, C-4). LCMS m/z calcd for C₁₇H₁₈ ⁷⁹BrNO₅S ([M]⁺) 427;found 428 ([M+H]⁺). HRMS m/z calcd for C₁₇H₁₈ ⁷⁹BrNNaO₅S 449.9987([M+Na]⁺); found 449.9977.

STEP G. Preparation of7-bromo-9-cyclopropyl-8-methoxy-9H-isothiazolo[5,4-B]quinoline-3,4-dione(17)(a) Anhydrous sodium hydrogen sulfide (Alfa Aesar, 53.3 mg, 0.95 mmol)is added in one portion to a solution of dimethylformamide (4.0 mL)containing 16 (158.1 mg, 0.37 mmol) at room temperature. The resultingsolution is heated at 50° C. for 1 h and allowed to cool to roomtemperature. The reaction mixture is quenched by addition of a 5%aqueous solution of hydrochloric acid (50 mL) and extracted with ethylacetate (100 mL). The organic extract is washed with brine (50 mL) andevaporated to dryness under reduced pressure to give crude ethyl7-bromo-1-cyclopropyl-2-mercapto-8-methoxy-4-oxo-1,4-dihydroquinoline-3-carboxylate.R_(f)˜0 (ethyl acetate). LCMS m/z calcd for C₁₆H₁₆ ⁷⁹BrNO₄S ([M]⁺) 397;found 398 ([M+H]⁺).

(b) A solution of sodium bicarbonate (316.9 mg, 3.77 mmol) in water (7.5mL) is added to a solution of ethyl7-bromo-1-cyclopropyl-2-mercapto-8-methoxy-4-oxo-1,4-dihydroquinoline-3-carboxylate(from above, ˜0.37 mmol) in tetrahydrofuran (7.5 mL) at roomtemperature. Hydroxylamine-O-sulfonic acid (214.7 mg, 1.90 mmol) isadded as a solid and in one portion to this mixture. The resulting ambersolution is stirred at room temperature for 2.5 h and quenched byaddition of an aqueous solution of 5% hydrochloric acid (50 mL). Thesolid that formed is collected by filtration, washed with an aqueoussolution of 5% hydrochloric acid (3×10 mL), washed with distilled water(3×10 mL), and dried in vacuo to give 17 as a tan solid. This productids of sufficient purity (≧95% by ¹H NMR spectroscopy) to use directlyin the next synthetic step. ¹H NMR (DMSO-d₆): δ1.00 (m, 2H, c-Pr CH₂),1.20 (m, 2H, c-Pr CH₂), 3.79 (s, 3H, OCH₃), 3.85 (m, 1H, c-Pr CH), 7.66(d, J=8.5 Hz, 1H, aromatic H-6), 7.93 (d, J=8.5 Hz, 1H, aromatic H-5),11.67 (br, 1H). ¹³C{¹H} NMR (DMSO-d₆): 811.5 (c-Pr CH₂), 35.1 (c-Pr CH),61.9 (OCH₃), 107.7, 122.9 (CH, C-5), 123.5 (C—Br, C-7), 127.9 (CH, C-6),128.0 (C-4a), 136.5 (C-8a), 146.6 (C—OCH₃), 164.5, 171.1 (C═O, C-4),171.2 (br). LCMS m/z calcd for C₁₄H₁₁ ⁷⁹BrN₂O₃S ([M]⁺) 366; found 367([M+H]⁺). HRMS m/z calcd for C₁₄H₁₁ ⁷⁹BrN₂NaO₃S 388.9571 ([M+Na]); found388.9577.

STEP H. General Method for the Preparation of9-cyclopropyl-8-methoxy-7-aryl-9H-isothiazolo[5,4-b]quinoline-3,4-diones(18)

Compound 18 is prepared by Suzuki Cross-Coupling Reaction of 17 and arylboronic acid (R═H) or aryl boronic ester (R=alkyl).

Under an atmosphere of argon, a reaction vessel is charged with 17 (0.1mmol), dimethylformamide (2 mL), tetrahydrofuran (2 mL),tetrakis(triphenylphosphine)palladium(0) (0.01-0.02 mmol), the desiredboronic acid or ester (0.3-0.4 mmol), and a 1 M aqueous solution ofsodium bicarbonate (1-2 mmol). The resulting mixture is irradiated withmicrowaves at 130° C. for 10-20 min, allowed to cool, and evaporated todryness under reduced pressure. The isolated residues are purified usingpreparative HPLC to give the desired products (95-99% purity). Thepurified products are isolated as TFA salts and converted to thecorresponding hydrochloride salts by addition of a 5% aqueous solutionof hydrochloric acid followed by evaporation; this process is repeatedtwice.

Example 3 Preparation of9-cyclopropyl-7-(2,6-dimethyl-pyridin-4-yl)-6-fluoro-9H-isothiazolo[5,4-B]quinoline-3,4-dione(XXIII) (25)

9-Cyclopropyl-7-(2,6-dimethyl-pyridin-4-yl)-6-fluoro-9H-isothiazolo[5,4-b]quinoline-3,4-dione(25) is prepared in accordance with the synthetic scheme set forthbelow:

STEP A. Preparation of 2,6-lutidine 1-oxide (20)

A solution of 2,6-lutidine (19, 23 ml, 200 mmol) and 50% hydrogenperoxide (15 ml) in glacial acetic acid (100 ml) is refluxed at 110° C.for 3 hours. The solution is then concentrated in vacuo at 40° C. toapproximately 60 ml. Water (20 ml) is added, and the concentrationprocess is repeated three times. The concentrated solution is furtherdried by lyophlizer overnight, yielding 26 g of 2,6-Lutidine 1-oxide(20) that contains approximately 10% acetic acid. ¹H NMR (300 MHz,CDCl₃) δ2.52 (s, 6H), 7.15 (m, 3H). MS, m/z 124 (M+1), 247 (2M+1).

STEP B. Preparation of 4-nitro-2, 6-lutidine 1-oxide (21)

A mixture of 2,6-lutidine 1-oxide (20, 15 g, 110 mmol) and concentratedsulfuric acid (98%, 30 ml) and concentrated nitric acid (70%, 12 ml) isheated under reflux for 3 hours. The mixture is poured into a largeexcess of ice and extracted with chloroform (3×100 ml). The combinedchloroform extracts are washed with aqueous sodium hydroxide and waterand dried over magnesium sulfate. Removal of the solvent yields pureyellow solid 4-Nitro-2, 6-lutidine 1-oxide (21). ¹H NMR (300 MHz, CDCl₃)δ2.64 (s, 6H), 8.08 (s, 2H). MS, m/z 169 (M+1), 210 (M+MeCN).

STEP C. Preparation of 4-amino-2, 6-lutidine (22)

A mixture of 4-Nitro-2,6-lutidine 1-oxide (21, 5.1 g, 30 mmol),palladized charcoal (10% Pd, 1 g) and acetic acid (2 ml) in methanol(200 ml) is hydrogenated under pressure (40 psi) over 10 hours using ahydrogenation apparatus. The reaction is followed with LC-MS. Afterfiltration, the filtrate is concentrated. The remaining oil is furtherdried by lyophilization, yielding 4.5 g of 4-amino-2, 6-lutidine (22)containing approximately 15% acetic acid. ¹H NMR (300 MHz, CDCl₃) δ2.3(s, 6H), 7.2 (s, 2H). MS, m/z 123 (M+1), 243 (2M−1).

STEP D. Preparation of 4-bromo-2, 6-lutidine (23)

Bromine (4 g) is added, with stirring over 10 minutes, to a mixture of4-amino-2, 6-lutidine (22, 1 g, approximately 6.5 mmol) in 48% HBr (12ml) at −10° C., followed by cooling to −20° C. A solution of sodiumnitrite (1.4 g) in water (4 ml) is added slowly. The mixture is stirredat −20° C. for 1 hour, and then warmed and left at room temperature for3 hours. The mixture is distilled. The oil fraction of the distillate isextracted with chloroform (3×10 ml). The combined extracts are driedover magnesium sulfate. After filtration, the filtrate is neutralized inan ice-bath using 2M butyl lithium in hexanes until the pH reaches 7. Alarge amount of salt forms. After filtration, the filtrate isconcentrated and dried, yielding 4-bromo-2,6-lutidine (23) oil. ¹H NMR(300 MHz, CDCl₃) δ2.82 (s, 6H), 7.5 (s, 2H). MS, m/z 186 and 188 (M+1).

STEP E. Preparation of 4-(2,6-dimethylpyridyl)boronic acid (24)

Butyl lithium (2M in hexanes, 0.6 ml, 1.2 mmol) is added to a solutionof 4-bromo-2, 6-lutidine (23, 0.2 g, 1 mmol) and triisopropyl borate(0.28 ml, 1.2 mmol) in solution of toluene (1.6 ml) and THF (0.4 ml)over 10 min at −40° C. under helium. The reaction is stirred at −40° C.for 30 minutes and then warmed to −20° C. 2N HCl (1 ml) is added toquench the reaction. The mixture is warmed to room temperature. Theaqueous layer is collected and neutralized using 5M NaOH. NaCl(approximately 0.4 g) is added. The aqueous layer is extracted with THF(3×10 ml), and the extracts are evaporated to dryness, yielding whitesolid 4-(2,6-dimethylpyridyl) boronic acid (24). ¹H NMR (300 MHz,DMSO-d₆) δ2.5 (s, 6H), 7.34 (s, 2H). MS, m/z 152 (M+1).

STEP F. Preparation of9-cyclopropyl-7-(2,6-dimethyl-pyridin-4-yl)-6-fluoro-9H-isothiazolo[5,4-B]quinoline-3,4-dione(25)

A mixture of tert-butyl7-bromo-9-cyclopropyl-6-fluoro-3,4-dioxoisothiazolo[5,4-b]quinoline-2(3H,4H,9H)-carboxylate(8, 22 mg, 0.048 mmol), 4-(2,6-Dimethylpyridyl)boronic acid (5, 23 mg,0.12 mmol) and Pd (PPh₃)₄ (4 mg) in a solution of DMF (1 ml) and 1MNaHCO₃ (0.22 ml) is sealed in a microwave reaction vessel with astirrer. After filling with helium, the mixture is microwaved at 100W,130° C. for 10 minutes. The reaction is followed by LC-MS. The reactionmixture is filtered and the filtrate evaporated to dryness. The residueis washed with a solution of methanol and ethyl ether (5:95) (3×3 ml),and dried in vacuum yielding pure product 25. ¹H NMR (300 MHz, DMSO-d₆)δ1.1-1.3 (m, 4H, —CH₂—), 1.53 (s, 1H, —CH—), 2.47 (s, 6H, —CH₃), 7.27(s, 2H, Ar), 7.93, (d, 1H, Ar), 8.2 (d, 1H, Ar). ¹⁹F NMR (350 MHz,DMSO-d₆) δ125 (s, 1F). MS, m/z 382 (M+1).

Example 4 Preparation of7-(1,2,3,6-tetrahydro-pyridin-4-yl)-9-cyclopropyl-6-fluoro-9H-isothiazolo[5,4B]-muinoline-3,4-dione(27)

STEP A. Preparation of1-acetyl-4-tributylstannyl-1,2,3,6-tetrahydropyridine

Caution: organotin compounds are toxic (Buck, B.; Mascioni, A.; Que, L,Jr., Veglia, G. J. Am. Chem. Soc. 2003, 125, 13316-13317, and referencescited therein). 1-(4-hydroxy-4-tributylstannanylpiperidin-1-yl)ethanoneis prepared using the two-step procedure described previously (Kiely, J.S.; Lesheski, L. E.; Schroeder, M. C. Preparation of Certain7-Substituted Quinolones. U.S. Pat. No. 4,945,160, Jul. 31, 1990).Because formation of1-(4-hydroxy-4-tributylstannanylpiperidin-1-yl)ethanone is reversible,this compound is used immediately after isolation (without purification)to generate 1-acetyl-4-tributylstannyl-1,2,3,6-tetrahydropyridine. Theisolated crude material is purified by flash column chromatography(eluting with 5% methanol in methylene chloride; R_(f) 0.30 (UVinactive)) to give the title product as a yellow oil. ¹H NMR (CDCl₃, 50°C.): δ0.89 (m, 15H, Bu), 1.33 (m, 6H, Bu), 1.50 (m, 6H, Bu), 2.07 (s,3H, NC(O)CH₃), 2.31 (m, 2H, H-2), 3.55 (m, 2H, H-3), 4.01 (m, 2H, H-6),5.76 (m, 1H, H-5). ¹H NMR spectroscopic data collected at roomtemperature matched those described in the literature, where twoconformational isomers were detected. LCMS m/z calcd for C₁₉H₃₇NO¹²⁰Sn([M]⁺) 415; found 416 ([M+H]⁺).

STEP B. Preparation of7-(1-acetyl-1,2,3,6-tetrahydro-pyridin-4-yl)-9-cyclopropyl-6-fluoro-9H-isothiazolo[5,4-B]quinoline-3,4-dione(26).

A mixture containing7-bromo-9-cyclopropyl-6-fluoro-9H-isothiazolo[5,4-b]quinoline-3,4-dione(7, 100.0 mg, 0.28 mmol),1-acetyl-4-tributylstannyl-1,2,3,6-tetrahydropyridine (190.0 mg, 0.46mmol), tetrakis(triphenylphosphine)palladium(0) (16.0 mg, 0.014 mmol),and dimethylformamide (6.0 mL) Is sparged with argon gas and irradiatedwith microwaves (5×10 min irradiations at 130° C.). (For cross-couplingexperiments described previously using C with conventional thermalheating, see: (a) Laborde, E.; Kiely, J. S.; Culbertson, T. P.;Lesheski, L. E. J. Med. Chem. 1993, 36, 1964-1970. (b) Kiely, J. S.;Laborde, E.; Lesheski, L. E.; Bucsh, R. A. J. Heterocyclic Chem. 1991,28, 1581-1585. (c) Laborde, E.; Kiely, J. S.; Lesheski, L. E.;Schroeder, M. C. J. Heterocyclic Chem. 1990, 28, 191-198) The resultingyellow solution is evaporated to dryness under reduced pressure (˜6 mmHg, 60° C.). The recovered solid is dissolved in a mixture containing10% methanol in methylene chloride (10 mL), precipitated via addition ofhexanes (100 mL), and collected by filtration. This process is repeatedonce. The product is purified further by flash column chromatography(eluting with 10% methanol in methylene chloride, R_(f) 0.50) to givepure 26 as a yellow solid (98% purity by HPLC). mp 243-244° C. ¹H NMR(CDCl₃/CD₃OD (12:1 v/v), 50° C.): δ1.32 (m, 2H, c-Pr CH₂), 1.45 (m, 2H,c-Pr CH₂), 2.18 (s, 3H, NC(O)CH₃), 2.65 (m, 2H, NCH₂CH₂), 3.45 (m, 1H,c-Pr CH), 3.73 (m, 1H, NCH₂CH₂), 3.87 (m, 1H, NCH₂CH₂), 4.23 (m, 1H,NCH₂CH), 4.31 (m, 1H, NCH₂CH), 6.19 (m, 1H, NCH₂CH), 7.88 (d,J_(H-F)=6.0 Hz, 1H, quinolone H-8), 8.08 (d, J_(H-F)=11.0 Hz, 1H,quinolone H-5). ¹⁹F{¹H} NMR (CDCl₃/CD₃OD (12:1 v/v), 50° C.): δ−119.4(s). NMR spectroscopic data collected at room temperature indicated thattwo conformational isomers were present. LCMS m/z calcd for C₂₀H₁₈FN₃O₃S([M]⁺) 399; found 400 ([M+H]⁺). HRMS m/z calcd for C₂₀H₁₈FN₃NaO₃S([M+Na]⁺) 422.0951; found 422.0951.

STEP C. Preparation of7-(1,2,3,6-tetrahydro-pyridin-4-yl)-9-cyclopropyl-6-fluoro-9H-isothiazolo[5,4-B]quinoline-3,4-dione(27)

Compound 26 (11.1 mg, 0.028 mmol) is dissolved partially in an aqueoussolution of hydrochloric acid (6 M, 3.0 mL) in air and heated at 90° C.to give an amber solution. After 22 h of heating, the solvent is removedunder reduced pressure. The residue is dissolved in water (˜3 mL) andtitrated to pH 7 with dilute sodium hydroxide. The precipitated solid iscollected by filtration, washed with water (2×10 mL), and dried in vacuoto give 27 as a yellow solid (98% purity by HPLC). mp >241-242° C. dec.¹H NMR (DMSO-d₆/acetic acid-d₄ (5:1 v/v)): δ1.19 (m, 2H, c-Pr CH₂), 1.29(m, 2H, c-Pr CH₂), 2.72 (m, 2H, NCH₂CH₂), 3.33 (m, 2H, NCH₂CH₂), 3.54(m, 1H, c-Pr CH), 3.80 (m, 2H, NCH₂CH), 6.21 (m, 1H, NCH₂CH), 7.87 (br,1H, aromatic), 7.91 (br, 1H, aromatic). ¹⁹F{¹H} NMR (DMSO-d₆/aceticacid-d₄ (5:1 v/v)): δ−121.4 (s). LCMS m/z calcd for C₁₈H₁₆FN₃O₂S ([M]⁺)357; found 358 ([M+H]⁺). FIRMS m/z calcd for C₁₈H₁₆FN₃NaO₂S ([M+Na]⁺)380.0845; found 380.0847.

Example 5 Preparation of(RAC)-7-(6-amino-5,6,7,8-tetrahydronaphthalen-2-yl)-9-cyclopropyl-6-fluoro-9H-isothiazolo[5,4-B]quinoline-3,4-dione(31)

STEP A. Preparation of(RAC)-tert-butyl(6-bromo-1,2,3,4-tetrahydronaphthalen-2-yl)carbamate(29).

(a) (rac)-6-Bromo-1,2,3,4-tetrahydronaphthalen-2-ylamine is preparedfrom 6-Bromo-3,4-dihydro-1H-naphthalen-2-one (28) via a generalprocedure described previously for the reductive amination of ketonesusing NaBH₃CN as reducing agent. (Borch, R. F.; Bernstein, M. D.; Durst,H. D. J. Am. Chem. Soc. 1971, 93, 2897-2904.). The purity of thismaterial (brown oil) was >95%, as determined by ¹H NMR spectroscopy, andwas used without further purification. ¹H NMR (CDCl₃): δ1.45 (br, 2H,NH₂), 1.56 (m, 1H, H-3), 1.98 (m, 1H, H-3), 2.47 (dd, J=16.0 Hz, 9.5 Hz,1H, H-1), 2.82 (m, 2H, H-4), 2.93 (dd, J=16.0 Hz, 4.5 Hz, 1H, H-1), 3.16(m, 1H, H-2), 6.92 (d, J=8.0 Hz, 1H, H-8), 7.22 (m, 2H, overlapping H-5and H-7). ¹³C NMR (CDCl₃): δ27.8 (CH₂, C-4), 32.5 (CH₂, C-3), 38.9 (CH₂,C-1), 47.0 (CH, C-2), 119.3 (C—Br, C-6), 128.7 (CH, C-7), 130.9 (CH,C-8), 131.4 (CH, C-5), 134.3 (C-8a), 138.2 (C-4a). LCMS m/z calcd forC₁₀H₁₂BrN ([M]⁺) 225; found 226 ([M+H]⁺).

(b) Di-tert-butyldicarbonate (575.7 mg, 2.64 mmol) in methylene chloride(5.0 mL) is added in one portion to a solution of methylene chloride(7.0 mL) at room temperature that contains(rac)-6-Bromo-1,2,3,4-tetrahydronaphthalen-2-ylamine (591.6 mg, 2.62mmol) and triethylamine (1.1 mL, 7.89 mmol). After stirring at roomtemperature for 19 h in air, the resulting amber solution is dilutedwith methylene chloride (25 mL), washed with a saturated aqueoussolution of sodium chloride (2×50 mL), dried over magnesium sulfate, andevaporated to dryness under reduced pressure to give the title compoundas a pale yellow solid. The purity of isolated 29 was >95%, asdetermined by ¹H NMR spectroscopy, and was used without further ° C. ¹HNMR (CDCl₃): δ1.45 (s, 9H, C(CH₃)₃), 1.70 (m, 1H, H-3), 2.04 (m, 1H,H-3), 2.55 (dd, J=16.5 Hz, 8.5 Hz, 1H, H-1), 2.84 (pseudo t, J=6.5 Hz,2H, H-4), 3.05 (dd, J=16.5 Hz, 5.0 Hz, 1H, H-1), 3.94 (br, 1H, H-2),4.58 (br, 1H, NH), 6.91 (d, J=8.0 Hz, 1H, H-8), 7.22 (m, 2H, overlappingH-5 and H-7). ¹³C NMR (CDCl₃): δ27.1 (CH₂, C-4), 28.4 (C(CH₃)₃), 28.7(CH₂, C-3), 35.6 (CH₂, C-1), 46.0 (CH, C-2), 79.4 (C(CH₃)₃), 119.6(C—Br, C-6), 128.9 (CH, C-7), 131.0 (CH, C-8), 131.5 (CH, C-5), 133.3(C-8a), 137.8 (C-4a), 155.3 (NHCO₂). LCMS m/z calcd for C₁₅H₂₀BrNO₂([M]⁺) 325; found 311 ([M−C₄H₇+CH₃CN]⁺, 22%), 270 ([M−C₄H₇]⁺, 81%), 267([M−C₅H₇O₂+CH₃CN]⁺, 43%), 226 ([M−C₅H₇O₂]+, 100%), 209 ([M−C₅H₁₀NO₂]⁺,94%).

STEP B. Preparation of(RAC)-tert-butyl[6-(4,4,5,5-tetramethyl[1,3,2]dioxaborolan-2-yl)-1,2,3,4-tetrahydronaphthalen-2-yl]carbamate(30)

The title compound, 30, is prepared via palladium-catalyzedcross-coupling reaction of 29 with bis(pinicolato)diboron using knownprocedures. (Ishiyama, T.; Murata, M.; Miyaura, N. J. Org. Chem. 1995,60, 7508-7510.) The crude product is purified by flash columnchromatography (eluting with 2% (v/v) methanol in methylene chloride;R_(f)0.41) to give pure(rac)-tert-butyl[6-(4,4,5,5-tetramethyl[1,3,2]dioxaborolan-2-yl)-1,2,3,4-tetrahydronaphthalen-2-yl]carbamateas an off-white, crystalline solid. mp 53-54° C. ¹H NMR (CDCl₃): δ1.33(s, 12H, OC(CH₃)₂C(CH₃)₂O), 1.45 (s, 9H, C(CH₃)₃), 1.73 (m, 1H, H-3),2.06 (m, 1H, H-3), 2.64 (dd, J=16.5 Hz, 8.0 Hz, 1H, H-1), 2.88 (pseudot, J=6.5 Hz, 2H, H-4), 3.12 (dd, J=16.5 Hz, 5.0 Hz, 1H, H-1), 3.97 (br,1H, H-2), 4.58 (br, 1H, NH), 7.07 (d, J=8.0 Hz, 1H, H-8), 7.54 (d, J=8.0Hz, 1H, H-7), 7.56 (s, 1H, H-5). ¹³C NMR (CDCl₃): δ24.8(OC(CH₃)₂C(CH₃)₂0), 26.9 (br, CH₂, C-4), 28.4 (C(CH₃)₃), 29.1 (br, CH₂,C-3), 36.3 (br, CH₂, C-1), 46.1 (br, CH, C-2), 79.3 (br, C(CH₃)₃), 83.7(OC(CH₃)₂C(CH₃)₂O), 126.3 (br, C—B, C-6), 129.0 (CH, C-8), 132.1 (CH,C-7), 134.9 (C-4a), 135.4 (CH, C-5), 137.8 (C-8a), 155.3 (NHCO₂). LCMSm/z calcd for C₂₁H₃₂BNO₄ ([M]⁺) 373; found 359 ([M−C₄H₇+CH₃CN]⁺, 21%),318 ([M−C₄14₇]⁺, 37%), 315 ([M−C₅H₇O₂+CH₃CN]⁺, 100%), 274 ([M−C₅H₇O₂]⁺,82%), 257 ([M−C₅H₁₀NO₂]⁺, 72%).

STEP C. Preparation of(RAC)-7-(6-amino-5,6,7,8-tetrahydronaphthalen-2-yl)-9-cyclopropyl-6-fluoro-9H-isothiazolo[5,4-B]quinoline-3,4-dione(31)

(a) A mixture containing7-bromo-9-cyclopropyl-6-fluoro-9H-isothiazolo[5,4-b]quinoline-3,4-dione(8) (31.8 mg, 0.090 mmol), 30 (68.0 mg, 0.182 mmol),tetrakis(triphenylphosphine)palladium(0) (7.2 mg, 0.006 mmol),dimethylformamide (1.5 mL), and a 1 M aqueous solution of sodiumbicarbonate (360 μL, 0.360 mmol) is sparged with argon gas andirradiated with microwaves (5-min irradiation at 120° C.). The resultinggreen, gelatinous mixture was filtered and evaporated to dryness underreduced pressure (˜6 mm Hg, 40° C.). The recovered residue (orange oil)is washed with diethyl ether (10 mL) to give a yellow solid. Thismaterial is dissolved in a mixture containing 25% methanol in chloroform(2.0 mL), precipitated via addition of diethyl ether (10 mL), andcollected by filtration; this process is repeated once. The product iswashed further with water (2×10 mL) and dried in vacuo to give(rac)-tert-Butyl[6-(9-cyclopropyl-6-fluoro-3,4-dioxo-2,3,4,9-tetrahydro-isothiazolo[5,4-b]quinolin-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl]carbamateas a yellow solid (93% purity by HPLC; the remaining material is9-cyclopropyl-6-fluoro-9H-isothiazolo[5,4-b]quinoline-3,4-dione). ¹H NMR(CDCl₃/CD₃OD (12:1 v/v), 50° C.): δ1.10 (m, 2H, c-Pr CH₂), 1.21 (m, 2H,c-Pr CH₂), 1.48 (s, 9H, C(CH₃)₃), 1.76 (m, 1H, H-3), 2.10 (m, 1H, H-3),2.68 (m, 1H, H-1), 2.86 (m, 2H, H-4), 3.08 (m, 1H, c-Pr CH), 3.12 (m,1H, H-1), 3.94 (m, 1H, H-2), 7.11 (m, 1H, aromatic), 7.20 (m, 2H,aromatic), 7.71 (m, 1H, aromatic), 7.92 (m, 1H, aromatic). ¹⁹F{¹H} NMR(CDCl₃/CD₃OD (12:1 v/v), 50° C.): δ−123.8 (s). NMR spectra collected atroom temperature contained broad, unresolved signals. LCMS m/z calcd forC₂₈H₂₈FN₃O₄S ([M]⁺) 521; found 522 ([M+H]⁺).

(b) In air, a solution of hydrogen chloride in acetic acid (1 M, 1.8 mL)is added at room temperature to a solution of methylene chloride (0.6mL) containing (rac)-tert-Butyl[6-(9-cyclopropyl-6-fluoro-3,4-dioxo-2,3,4,9-tetrahydro-isothiazolo[5,4-b]quinolin-7-yl)-1,2,3,4-tetrahydronaphthalen-2-yl]carbamate (11.4mg, 0.022 mmol). A yellow precipitate begins to appear within minutes ofaddition of the solution of hydrogen chloride. After stirring themixture at room temperature for 18 h, additional methylene chloride isadded (2 mL). The precipitate is collected by filtration, washed withmethylene chloride (4×5 mL), and dried in vacuo to give pure 31 (97%purity by HPLC analysis) as a yellow powder. mp >257-258° C. dec. ¹H NMR(DMSO-d₆, 60° C.): δ1.21 (m, 2H, c-Pr CH₂), 1.29 (m, 2H, c-Pr CH₂), 1.80(m, 1H, H-3), 2.12 (m, 1H, H-3), 2.83 (dd, J=17.0 Hz, 10.0 Hz, 1H, H-1),2.90 (m, 2H, H-4), 3.13 (dd, J=17.0 Hz, 6.0 Hz, 1H, H-1), 3.46 (m, 1H,H-2), 3.57 (m, 1H, c-Pr CH), 7.25 (d, J=8.0 Hz, 1H, H-8), 7.37 (s, 1H,H-5), 7.39 (d, J=8.0 Hz, 1H, H-7), 7.93 (d, J_(H-F)=10.5 Hz, 1H, ITQH-5), 8.01 (d, J_(H-F)=6.5 Hz, 1H, ITQ H-8). ¹⁹F{¹1H} NMR (DMSO-d₆, 60°C.): δ−123.3 (s). NMR spectra collected at room temperature containedbroad, unresolved signals. LCMS m/z calcd for C₂₃H₂₀FN₃O₂S ([M]⁺) 421;found 422 ([M+H]⁺). HRMS m/z calcd for C₂₃H₂₀FN₃Na0₂S ([M+Na]⁺)444.1158; found 444.1152.

Example 6 Preparation of9-cyclopropyl-6-fluoro-7-(2-(Pyridin-2-yl)ethynyl)isothiazolo[5,4-B]Quinoline-3,4(2H,9H)-dione (Compound 60)

Pd (PPh₃)₄ (4.5 mg) is added to a stirred solution of7-bromo-9-cyclopropyl-6-fluoro-9H-isothiazolo[5,4-b]quinoline-3,4-dione(23 mg, 0.065 mmole) in DMF (1mL), followed by the addition of2-ethynylpyridine (2 equivalents, 0.13 mmole) and diisopropyl amine(0.15 mL.) under argon at room temperature. The reaction tube is sealedand then stirred in a microwave (100W, 90° C.) until complete asmonitored by LC/MS. The reaction mixture is filtered and the filtrateconcentrated in vacuo. The residue is dissolved in a mixture of DMF:CHCl₃: MeOH (1:8:1) and precipitated by adding diethyl ether (1-2 ml).This process is repeated three to five times. The resulting solid,9-cyclopropyl-6-fluoro-7-(2-(pyridin-2-yl)ethynyl)isothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione, is washed with water toremove salt. The product is dried and analyzed. HPLC may be needed toobtain adequately pure product.

Example 7 Preparation of9-cyclopropyl-7-substituted-6-fluoro-9H-isoxazolo[5,4-B]quinoline-3,4-diones

A solution of a1-cyclopropyl-7-substituted-6-fluoro-2-methanesulfinyl-4-oxo-1,4-dihydro-quinoline-3-carboxylicacid ethyl ester (X) (10 mg, 0.023 mmol), hydroxyurea (3 mg, 0.039 mmol)and 1,8-diazabicyclo[5,4,0]undec-7-ene (DBU) (6 μl, 0.04 mmol) inmethanol (5 ml) is stirred overnight at room temperature. The mixture isevaporated to dryness in vacuo. The resulting residue was washed with 2%acetic acid. The remaining solid(9-Cyclopropyl-7-substituted-6-fluoro-9H-isoxazolo[5,4-b]quinoline-3,4-dione,(Y) is collected and dried in vacuo.

Example 8 Procedure for N- and O-Alkylation of Isothiazoloquinolones

Cesium carbonate (0.25 mmol) and akyl halide (0.10-0.50 mmol) are addedsequentially to a solution of isothiazoloquinolone (A) (0.10 mmol) indimethylformamide (20 mL) at room temperature. The resulting mixture isstirred for 18 h. The reaction mixture is quenched with water (100 mL),and the product extracted with ethyl acetate (3×150 mL). The combinedorganic extracts are washed with brine (100 mL), dried over magnesiumsulfate, and evaporated under reduced pressure to give mixtures of thedesired O- (B, major) and N-alkylated (C, minor) isothiazoquinolones.The mixture is separated into the individual O- and N-alkylated productsby column chromatography.

Example 9 Additional compounds of Formula I and Formula II

The following compounds, shown in Table I, are made by the methodsdisclosed in Examples 1 to 8. Those of ordinary skill in the art willrecognize that the procedures and starting materials may be varied inorder to obtain the compounds disclosed herein.

TABLE I 19F- No. Structure Name MS 1H-NMR NMR 33

9-cyclopropyl-8- methoxy-7-(4- (piperidin-1- ylmethyl) phenyl)isothiazolo[5,4- b]quinoline- 3,4(2H,9H)- dione LCMS (APCI): m/z calcdfor C₂₆H₂₇N₃O₃S [M⁺] 461.58; found ([M + H]⁺) 462.10 ¹H NMR (DMSO- d₆):δ 1.07 (m, 2H), 1.22 (m, 2H), 1.80 (m, 6H), 2.87 (m, 2H), 3.29 (m, 2H),3.35 (s, 3H), 3.85 (m, 1H), 7.42 (d, J = 8.4 Hz, 1H), 7.71 (d, J = 8.4Hz, 2H), 7.76 (d, J = 8.4 Hz, 2H), 8.06 (d, J = 8.4 Hz, 1H) 34

9-cyclopropyl-6- fluoro- 8-methoxy- 7-(2- methylpyridin- 4-yl)isothiazolo[5,4- b]quinoline- 3,4(2H,9H)- dione LCMS (APCI): m/z calcdfor C₂₀H₁₆FN₃O₃S [M⁺] 397.42; found ([M + H]⁺) 398.54 ¹H NMR (DMSO- d₆):δ 1.01 (m, 2H), 1.10 (m, 2H), 2.50 (s, 3H), 3.37 (s, 3H), 3.76 (m, 1H),7.28 (d, J_(H-H) = 5.0 Hz, 1H), 7.35 (s, 1H), 7.75 (d, J_(H-F) = 9.5 Hz,1H), 8.55 (d, J_(H-H) = 5.0 Hz, 1H) ¹⁹F NMR (DMSO- d₆): δ −119.0 (s) 35

9-cyclopropyl-8- methoxy-7- (1,2,3,4- tetrahydro- isoquinolin-6-yl)isothiazolo [5,4- b]quinoline- 3,4(2H,9H)- dione LCMS (APCI): m/zcalcd for C₂₃H₂₁N₃O₃S [M⁺] 419.50; found ([M + H]⁺) 420.54 ¹H NMR (DMSO-d₆): δ 1.06 (m, 2H), 1.22 (m, 2H), 3.10 (m, 2H), 3.36 (s, 3H), 3.43 (m,2H), 3.85 (m, 1H), 4.35 (m, 2H), 7.36 (d, J = 8.0 Hz, 1H), 7.40 (d, J =8.5 Hz, 1H), 7.50 (m, 1H), 7.56 (m, 1H), 8.05 (d, J = 8.5 Hz, 1H) 36

9-cyclopropyl- 6-fluoro- 8-methoxy- 7-(6- methylpyridin-3-yl)isothiazolo [5,4- b]quinoline- 3,4(2H,9H)- dione LCMS (APCI): m/zcalcd for C₂₀H₁₆FN₃O₃S [M⁺] 397.42; found ([M + H]⁺) 398.07 ¹H NMR(DMSO- d₆): δ 1.15 (m, 4H), 2.74 (s, 3H), 3.43 (s, 3H), 3.84 (m, 1H),7.86 (m, 2H), 8.36 (d, J_(H,F) = 9.1 Hz, 1H), 8.80 (s, br, 1H) ¹⁹F NMR(DMSO- d₆): δ −119.2 (s) 37

9-cyclopropyl- 6-fIuoro- 8-methoxy- 7-((R)-1- methylisoindolin- 5-yl)isothiazolo [5,4- b]quinoline- 3,4(2H,9H)- dione LCMS (APCI): m/zcalcd for C₂₃H₂₀FN₃O₃S [M⁺] 437.49; found ([M + H]⁺) 438.49 ¹H NMR(DMSO- d₆): δ 1.06 (m, 2H), 1.18 (m, 2H), 1.65 (d, J = 7.0 Hz, 3H), 3.39(s, 3H), 3.81 (m, 1H), 4.48-4.67 (m, 2H), 5.00 (m, 1H),7.55 (m, 3H),7.79 (d, J = 9.5 Hz, 1H) ¹⁹F NMR (DMSO- d₆): δ −118.7 (s) 38

9-cyclopropyl- 6-fluoro- 7-(isoindolin- 5-yl)-8- methoxy-isothiazolo[5,4- b]quinoline- 3,4(2H,9H)- dione LCMS (APCI): m/z calcdfor C₂₂H₁₈FN₃O₃S [M⁺] 423.20; found ([M + H]⁺) 424.17 ¹H NMR (DMSO- d₆):δ 1.06 (m, 2H), 1.18 (m, 2H), 3.39 (s, 3H), 3.82 (m, 1H), 4.59 (m, 4H),7.49-7.60 (m, 3H), 7.79 (d, J = 9.5 Hz, 1H) ¹⁹F NMR (DMSO- d₆): δ −118.7(s) 39

9-cyclopropyl- 6-fIuoro- 8-methoxy- 7-(1,2,3,4- tetrahydro-isoquinolin-6- yl)isothiazolo [5,4- b]quinoline- 3,4(2H,9H)- dione LCMSm/z calcd for C₂₃H₂₀FN₃O₃S ([M⁺]) 437; found 437 ([M + H]⁺) ¹H NMR(DMSO- d₆): δ 1.06 (m, 2H), 1.18 (m, 2H), 3.09 (m, 2H), 3.40 (s, 3H),3.43 (m, 2H), 3.81 (m, 1H), 4.35 (m, 2H), 7.38 (m, 3H), 7.78 (d, J = 9.5Hz, 1H) ¹⁹F NMR (DMSO- d₆): δ −118.7 (s) 40

9-cyclopropyl-8- methoxy-7- (2-methyl- 1,2,3,4- tetrahydro-isoquinolin-6- yl)isothiazolo [5,4- b]quinoline- 3,4(2H,9H)- dione LCMS(APCI): m/z calcd for C₂₄H₂₃N₃O₃S ([M]⁺) 433; found 434 ([M + H]⁺) ¹HNMR (DMSO- d₆ 80° C.): δ 1.07 (m, 2H), 1.25 (m, 2H), 2.97 (s, 3H), 3.21(m, 2H), 3.42 (s, 3H), 3.54 (m, 2H), 3.89 (m, 1H), 4.44 (m, 2H), 7.33(m, 1H), 7.41 (m, 1H), 7.49-7.60 (m, 2H), 8.09 (m, 1H). 41

9-cyclopropyl- 6-fluoro- 8-methoxy-7- (2-methyl- 1,2,3,4- tetrahydro-isoquinolin-6- yl)isothiazolo [5,4- b]quinoline- 3,4(2H,9H)- dione LCMS(APCI): m/z calcd for C₂₄H₂₂FN₃O₃S ([M]⁺) 451; found 452 ([M + H]⁺) ¹HNMR (DMSO- d₆, 80° C.): δ 1.08 (m, 2H), 1.22 (m, 2H), 2.98 (s, 3H), 3.20(m, 2H), 3.45 (s, 3H), 3.55 (m, 2H), 3.85 (m, 1H), 4.46 (m, 2H),7.34-7.45 (m, 3H), 7.81 (d, J_(H-F) = 9.5 Hz, 1H). ¹⁹F NMR (DMSO- d₆,80° C.): δ −117.8 (s). 42

7-(3-amino-4- fluorophenyl)-9- cyclopropyl-8- methoxy- isothiazolo[5,4-b]quinoline- 3,4(2H,9H)- dione LCMS (APCI): m/z calcd for C₂₀H₁₆FN₃O₃S[M⁺] 397.42; found ([M + H]⁺) 398.10 ¹H NMR (DMSO- d₆): δ 1.12 (m, 4H),3.37 (s, 3H), 3.84 (m, 1H), 6.83 (m, 1H), 7.12 (m, 2H), 7.33 (d, J = 8.2Hz, 1H), 8.01 (d, J = 8.2 Hz, 1H) ¹⁹F NMR (DMSO- d₆): δ −135.4 (s) 43

9-cyclopropyl-8- methoxy-7-(2- methylpyridin-4- yl)isothiazolo [5,4-b]quinoline- 3,4(2H,9H)- dione LCMS (APCI): m/z calcd for C₂₀H₁₇N₃O₃S[M⁺] 379.43; found ([M + H]⁺) 380.08 ¹H NMR (DMSO- d₆): δ 1.16 (m, 4H),3.44 (s, 3H), 3.86 (m, 1H), 7.54 (d, J = 8.1 Hz, 1H), 7.96 (d, J = 5.9Hz, 1H), 8.02 (s, br, 1H), 8.13 (d, J = 8.1 Hz, 1H), 8.81 (d, J = 5.9Hz, 1H) 44

(E)-4-(9- cyclopropyl-6- fluoro-3,4- dioxo- 2,3,4,9- tetrahydro-isothiazolo[5, 4-b]quinolin-7- yl)picolinalde- hyde O- methyl oxime LCMSm/z calcd for C₂₀H₁₅FN₄O₃S ([M]⁺) 410; found 411 ([M + H]⁺) ¹H NMR(DMSO- d₆): δ 1.29 (m, 2H), 1.40 (m, 2H), 3.68 (m, 1H), 4.00 (s, 3H),7.80 (m, 1H), 8.09 (m, 2H), 8.24 (d, J = 6.3 Hz, 1H), 8.32 (s, 1H), 8.83( d, J = 5.2 Hz, 1H). ¹⁹F NMR (DMSO- d₆): δ −123.7 (s). 45

9-cyclopropyl- 6-fluoro- 7-(2- (hydroxymethyl) pyridin- 4-yl)isothiazolo[5,4- b]quinoline- 3,4(2H,9H)- dione LCMS m/z calcd for C₁₉H₁₄FN₃O₃S([M]⁺) 383; found 384 ([M + H]⁺) ¹H NMR (DMSO- d₆): δ 1.21 (m, 2H), 1.31(m, 2H), 3.59 (m, 1H), 4.73 (s, 2H), 4.36 (bs, 2H), 7.77 (d, J = 5.3 Hz,1H), 7.91 (m, 1H), 8.01 (d, J = 10.7 Hz, 1H), 8.16 (d, J = 6.2 Hz, 1H),8.72 (d, J = 5.3 Hz, 1H). ¹⁹F NMR (DMSO- d₆): δ −123.5 (s). 46

9-cyclopropyl- 6-fluoro- 7-(6-fluoropyridin- 3-yl)- 8-methoxy-isothiazolo [5,4-b] quinoline- 3,4(2H,9H)- dione LCMS (APCI): m/z calcdfor C₁₉H₁₃F₂N₃O₃S [M⁺] 401.39; found ([M + H]⁺) 402.03 ¹H NMR (DMSO-d₆): δ 1.14 (m, 4H), 3.39 (s, 3H), 3.82 (m, 1H), 7.40 (dd, J₁ = 8.3 Hz,J₂ = 2.7 Hz, 1H), 7.83 (d, J_(H,F) = 9.8 Hz, 1H), 8.21 (t, J = 8.3 Hz,1H), 8.83 (s, br, 1H) 19F NMR (DMSO- d₆): δ −119.3 (s), −69.14 (s) 47

9-cyclopropyl- 6-fluoro- 7-(2-fluoropyridin- 3-yl)- 8-methoxy-isothiazolo [5,4-b] quinoline- 3,4(2H,9H)- dione LCMS (APCI): m/z calcdfor C₁₉H₁₃F₂N₃O₃S [M⁺] 401.39; found ([M + H]⁺) 402.02 ¹H NMR (DMSO-d₆): δ 1.12 (m, 4H), 3.39 (s, 3H), 3.82 (m, 1H), 7.40 (dd, J₁ = 8.2 Hz,J₂ = 2.7 Hz, 1H), 7.83 (d, J_(H,F) = 9.9 Hz, 1H), 8.21 (t, J = 8.2 Hz,1H), 8.43 (s, br, 1H) ¹⁹F NMR (DMSO- d₆): δ −119.0 (s), −70.1 (s) 48

9-cyclopropyl- 6-fIuoro- 7-(2-fluoropyridin- 4-yl)- 8-methoxy-isothiazolo [5,4-b] quinoline- 3,4(2H,9H)- dione LCMS (APCI): m/z calcdfor C₁₉H₁₃F₂N₃O₃S [M⁺] 401.39; found ([M + H]⁺) 402.00 ¹H NMR (DMSO-d₆): δ 1.14 (m, 4H), 3.45 (s, 3H), 3.82 (m, 1H), 7.43 (s, 1H), 7.55 (d,J = 5.1 Hz, 1H), 7.83 (d, J_(H,F) = 9.6 Hz, 1H), 8.43 (d, J = 5.1 Hz,1H) ¹⁹F NMR (DMSO- d₆): δ −120.0 (s), −69.2 (s) 49

9-cyclopropyl- 7-(4- (hydroxymethyl) phenyl)- 8-methoxy- isothiazolo[5,4-b] quinoline- 3,4(2H,9H)- dione LCMS m/z calcd for C₂₁H₁₈N₂O₄S([M]⁺) 394; found [M + H] :¹H-NMR (DMSO- D₆): δ 1.072-1.096 (m, 2H),1.222- 1.269 (m, 2H), 3.343 (s, 3H), 3.828-3.899 (m, 1H), 4.586 (s, 2H),7.423 (d, J = 8.1 Hz, 1H), 7.464 (d, J = 8.4 Hz, 2H), 7.626 (d, J = 8.1Hz, 2H), 8.056 (d, J = 8.1 Hz, 1H) 50

9-cyclopropyl- 7-(3- hydroxy- phenyl)-8- methoxy- isothiazolo[5,4-b]quinoline- 3,4(2H,9H)- dione LCMS m/z calcd for C₂₀H₁₆N₂O₄S ([M]⁺)380; found [M + H] ¹H-NMR (DMSO- D₆): δ 1.058- 1.068 (m, 2H),1.213-1.234 (m, 2H), 3.372 (s, 3H), 3.842-3.899 (m, 1H), 6.828 (dd, J =1.5, 0.9 Hz, 1H), 7.039- 7.062 (m, 2H), 7.311 (t, J = 8.1 Hz, 1H), 7.377(d, 8.4 Hz, 1H), 8.036 (d, J = 8.1 Hz, 1H), 9.575 (brs, 1H) 51

7-(4-amino- phenyl)-9- cyclopropyl- 8- methoxy- isothiazolo[5,4-b]quinoline- 3,4(2H,9H)- dione LCMS m/z calcd for C₂₀H₁₇N₃O₃S ([M]⁺)379; found [M + H] ¹H-NMR (DMSO- D₆): δ 1.031- 1.084 (m, 2H),1.192-1.234 (m, 2H), 3.351 (s, 3H), 3.826-3.898 (m, 1H), 7.101 (brs,2H), 7.405 (d, J = 8.4 Hz, 1H), 7.596(d, J = 7.5 Hz, 2H), 8.032 (d, J =8.4 Hz, 1H). 52

7-(4-amino- 3,5- dimethylphenyl)- 9- cyclopropyl-6- fluoro-8- methoxy-isothiazolo[5,4- b]quinoline- 3,4(2H,9H)- dione LCMS (APCI): m/z calcdfor C₂₂H₂₀FN₃O₃S [M⁺] 425.48; found ([M + H]⁺) 426.00 ¹H NMR (DMSO- d₆):δ 1.10 (m, 4H), 2.31 (s, 6H), 3.37 (s, 3H), 3.82 (m, 1H), 7.19 (s, 2H),7.74 (d, J_(H,F) = 10.0 Hz, 1H) ¹⁹F NMR (DMSO- d₆): δ −118.4 (s) 53

7-(4-amino- 3,5- dimethylphenyl)- 9- cyclopropyl-6- fluoro-isothiazolo[5,4- b]quinoline- 3,4(2H,9H)- dione LCMS (APCI): m/z calcdfor C₂₁H₁₈FN₃O₂S [M⁺] 395.46; found ([M + H]⁺) 397.20 ¹H NMR (DMSO- d₆):δ 1.29 (m, 4H), 2.28 (s, 6H), 3.82 (m, 1H), 7.28 (s, 2H), 7.90 (d,J_(H,F) = 11.3 Hz, 1H), 8.01 (d, J = 5.9 Hz, 1H) ¹⁹F NMR (DMSO- d₆): δ−123.3 (s) 54

9-cyclopropyl- 6-fluoro- 7-(4- (hydroxymethyl) phenyl)-8- methoxyisothiazolo [5,4-b] quinoline- 3,4(2H,9H)- dione LCMS m/z calcd forC₂₁H₁₇FN₂O₄S ([M]⁺) 412; found [M + H] ¹H-NMR (DMSO- D₆): δ 1.068- 1.096(m, 2H), 1.166-1.207 (m, 2H), 3.367 (s, 3H), 3.794-3.865 (m, 1H), 4.596(s, 2H), 7.488 (s, 4H), 7.789 (d, J = 9.3 Hz, 1H) ¹⁹F (DMSO- D₆): δ−118.79 55

9-cyclopropyl- 7-(2- fluoropyridin- 4-yl)-8- methoxy- isothiazolo[5,4-b]quinoline- 3,4(2H,9H)- dione LCMS m/z calcd for C₁₉H₁₄FN₃O₃S ([M]⁺)383; found [M + H] ¹H-NMR (DMSO- D₆): δ 1.091- 1.115 (m, 2H),1.202-1.244 (m, 2H), 3.745 (s, 3H), 3.854-3.879 (m, 1H), 7.472 (s, 1H),7.528 (d, J = 8.1 Hz, 1H), 7.656 (dt, J = 5.1, 1.5 Hz, 1H), 8.108 (d, J= 8.4 Hz, 1H), 8.401 (d, J = 5.1 Hz, 1H) ¹⁹F (DMSO- D₆): δ −69.01 56

9-cyclopropyl- 6-fluoro- 7-(2-methyl- pyridin-4- yl)isothiazolo [5,4-b][1,8] naphthyridine- 3,4(2H,9H)- dione LCMS m/z calcd for C₁₈H₁₃FN₄O₂S([M]⁺) 368; found 369 ([M + H]⁺) ¹H NMR (DMSO- d₆): δ 1.21 (m, 2H), 1.30(m, 2H), 2.75 (s, 3H), 3.48 (m, 1H), 8.25 (m, 1H), 8.29 (m, 1H), 8.51(d, J = 10.5 Hz, 1H), 8.85 (d, J = 6.0 Hz, 1H) ¹⁹F NMR (DMSO- d₆): δ−127.4 (s) 57

9-cyclopropyl- 7-(2,6- dimethylpyridin- 4-yl)-6- fluoro-isothiazolo[5,4- b][1,8] naphthyridine- 3,4(2H,9H)- dione LCMS m/z calcdfor C₁₉H₁₅FN₄O₂S ([M]⁺) 382; found 383 ([M + H]⁺) ¹H NMR (DMSO- d₆): δ1.21 (m, 2H), 1.31 (m, 2H), 2.73 (s, 6H), 3.47 (m, 1H), 8.15 (m, 2H),8.52 (d, J = 10.5 Hz, 1H) ¹⁹F NMR (DMSO- d₆): δ −127.1 (br) 58

9-cyclopropyl-8- methoxy-7- (pyridin-3- yl)isothiazolo [5,4-b]quinoline- 3,4(2H,9H)- dione LCMS m/z calcd for C₁₉H₁₅N₃O₃S ([M]⁺)365; found M⁺ ¹H-NMR (DMSO- D₆): δ 1.112- 1.121 (m, 2H), 1.206-1.248 (m,2H), 3.391 (s, 3H), 3.834-3.905 (m, 1H), 7.546 (d, J = 8.1 Hz, 1H),7.825 (dd, J = 2.4, 5.1 Hz, 1H), 8.129 (d, J = 8.1 Hz, 1H), 8.391 (d, J= 7.8 Hz, 1H), 8.796 (d, J = 4.5 Hz, 1H), 8.999 (brs, 1H). 59

9-cyclopropyl-7- (isoquinolin- 6-yl)-8- methoxy- isothiazolo[5,4-b]quinoline- 3,4(2H,9H)- dione LCMS m/z calcd for C₂₃H₁₇N₃O₃S ([M]⁺)415, found M⁺ ¹H-NMR (DMSO- D₆): δ 1.105- 1.158 (2H, m), 1.237-1.278(2H, m), 3.369 (3H, s), 3.860- 3.918 (1H, m), 7.601 (1H, d, J = 8.1 Hz),7.936 (1H, dd, J = 3.6, 4.8 Hz), 8.151 (1H, d, J = 8.4 Hz), 8.271 (1H,dd, J = 6.9, 1.8 Hz), 8.383 (1H, d, J = 8.7 Hz), 8.492 (1H, d, J = 1.8Hz), 8.958 (1H, d, J = 8.1 Hz), 9.208 (1H, dd, J = 3.6, 1.2 Hz). 60

5-(9- cyclopropyl-6- fluoro-3,4- dioxo- 2,3,4,9- tetrahydro-isothiazolo[5, 4-b]quinolin-7- yl) nicotinonitrile LCMS (APCI): m/zcalcd for C₁₉H₁₁FN₄O₂S [M⁺] 378.38; found ([M + H]⁺) 378.95 ¹H NMR(DMSO- d₆): δ 1.31 (m, 4H), 3.61 (m, 1H), 8.02 (d, J_(H,F) = 10.5 Hz,1H), 8.24 (d, J = 5.2 Hz, 1H), 8.70 (s, br, 1H), 9.16 (s, br, 2H) ¹⁹ FNMR (DMSO- d₆): δ −124.1 (s) 61

5-(9- cyclopropyl-8- methoxy-3,4- dioxo- 2,3,4,9- tetrahydro-isothiazolo[5, 4-b]quinolin-7- yl) nicotinonitrile LCMS (APCI): m/zcalcd for C₂₀H₁₄N₄O₃S [M⁺] 390.42; found ([M + H]⁺) 392.09 ¹H NMR (DMSO-d₆): δ 1.14 (m, 4H), 3.39 (s, 3H), 3.84 (m, 1H), 7.54 (d, J = 8.2 Hz,1H), 8.10 (d, J = 8.2 Hz, 1H), 8.60 (m, 1H), 9.09 (d, J = 2.2 Hz, 1H),9.12 (s, J = 2.2 Hz, 1H) 62

5-(9- cyclopropyl-6- fluoro-8- methoxy-3,4- dioxo-2,3,4,9- tetrahydro-isothiazolo[5, 4-b]quinolin-7- yl) nicotinonitrile LCMS (APCI): m/zcalcd for C₂₀H₁₃FN₄O₃S [M⁺] 408.41; found ([M + H]⁺) 409.15 ¹H NMR(DMSO- d₆): δ 1.16 (m, 4H), 3.39 (s, 3H), 3.84 (m, 1H), 7.86 (d, J_(H.F)= 9.2 Hz, 1H), 8.61 (s, 1H), 9.04 (s, 1H), 9.15 (s, 1H) ¹⁹F NMR (DMSO-d₆): δ −119.4 (s) 63

9-cyclopropyl- 7-(6- fluoro-2- methylpyridin- 3-yl)-8- methoxyisothiazolo [5,4-b] quinoline- 3,4(2H,9H)- dione LCMS m/z calcd forC₂₀H₁₆FN₃O₃S ([M]⁺) 397; found [M + H] ¹H-NMR (DMSO- D₆): δ 1.043- 1.051(2H, m), 1.195-1.213 (2H, m), 2.375 (3H, s), 3.344 (3H, s), 3.813- 3.885(1H, m), 7.156 (1H, dd, J = 5.1, 3.0 Hz), 7.327 (1H, d, J = 8.4 Hz),7.950 (1H, t, J = 8.4 Hz), 8.093 (1H, d, J = 8.1 Hz) ¹⁹F (DMSO- D₆): δ−70.31. 64

9-cyclopropyl- 6-fluoro- 7-(6-fluoro-2- methylpyridin-3- yl)isothiazolo[5,4- b]quinoline- 3,4(2H,9H)- dione LCMS m/z calcd for C₁₉H₁₃F₂N₃O₂S([M]⁺) 385; found M⁺ ¹H-NMR (DMSO- D₆): δ 1.246- 1.288 (2H, m),1.333-1.353 (2H, m), 2.377 (3H, s), 3.395 (3H, s), 3.373- 3.746 (1H, m),7.211 (1H, dd, J = 5.1, 3.0 Hz), 7.973-8.051 (3H, m). ¹⁹F (DMSO- D₆): δ−68.94, −121.54. 65

9-cyclopropyl- 7-(2,6- dimethylpyridin- 3-yl)-8- methoxy-isothiazolo[5,4- b]quinoline- 3,4(2H,9H)- dione LCMS m/z calcd forC₂₁H₁₉N₃O₃S ([M]⁺) 393; found [M + H] ¹H-NMR (DMSO- D₆): δ 1.068- 1.078(2H, m), 1.198-1.217 (2H, m), 2.608 (3H, s), 2.767 (3H, s), 3.378 (3H,s), 3.820- 3.877 (1H, m), 7.372 (1H, d, J = 9.0 Hz), 7.791 (1H, brs),8.143 (1H, d, J = 8.7 Hz), 8.319 (1H, brs). 66

9-cyclopropyl- 6-fluoro- 7-(pyridin-4- yl)isothiazolo [5,4-b][1,8]naphthyridine- 3,4(2H,9H)- dione LCMS m/z calcd for C₁₇H₁₁FN₄O₂S ([M]⁺)354; found 355 ([M + H]⁺) ¹H NMR (DMSO- d₆): δ 1.27 (m, 2H), 1.35 (m,2H), 3.54 (m, 1H), 8.26 (m, 2H), 8.52 (d, J = 10.5 Hz, 1H), 8.93 (m, 2H)¹⁹F NMR (DMSO- d₆): δ −128.2 (s) 67

9-cyclopropyl- 6-fIuoro- 7-(6- methylpyridin-3- yl)isothiazolo[5,4-b][1,8] naphthyridine- 3,4(2H,9H)- dione LCMS m/z calcd forC₁₈H₁₃FN₄O₂S ([M]⁺) 368; found 369 ([M + H]⁺) ¹H NMR (DMSO- d₆): δ 1.20(m, 2H), 1.27 (m, 2H), 2.68 (s, 3H), 3.48 (m, 1H), 7.83 (m, 1H), 8.45(d, J = 10.5 Hz, 1H), 8.74 (m, 1H), 9.19 (m, 1H) ¹⁹F{¹H} NMR (DMSO- d₆):δ −129.1 (s) 68

9-cyclopropyl- 6-fluoro- 7-(pyridin-3- yl)isothiazolo [5,4-b][1,8]naphthyridine- 3,4(2H,9H)- dione LCMS m/z calcd for C₁₇H₁₁FN₄O₂S ([M]⁺)354; found 355 ([M + H]⁺) ¹H NMR (DMSO- d₆): δ 1.22 (m, 2H), 1.28 (m,2H), 3.48 (m, 1H), 7.70 (dd, J = 7.5 Hz, J = 5.0 Hz, 1H), 8.42 (d, J =10.5 Hz, 1H), 8.55 (m, 1H), 8.76 (m, 1H), 9.27 (m, 1H) ¹⁹F{¹H} NMR(DMSO- d₆): δ −129.4 (s) 69

7-(4- (aminomethyl) phenyl)- 9-cyclopropyl- 6-fluoro- isothiazolo[5,4-b][1,8] naphthyridine- 3,4(2H,9H)- dione LCMS m/z calcd for C₁₉H₁₅FN₄O₂S([M]⁺) 382; found 383 ([M + H]⁺, 70%), 366 (100%) ¹H NMR (DMSO- d₆): δ1.15- 1.31 (m, 4H), 3.49 (m, 1H), 4.08 (q, J = 6.0 Hz, 2H), 7.65 (m,2H), 8.11 (m, 2H), 8.36 (d, J = 10.5 Hz, 1H) ¹⁹F{¹H} NMR (DMSO- d₆): δ−129.3 (s) 70

9-cyclopropyl- 7-(2,6- difluoropyridin- 3-yl)-8- methoxy-isothiazolo[5,4- b]quinoline- 3,4(2H,9H)- dione LCMS m/z calcd forC₁₉H₁₃F₂N₃O₃S ([M]⁺) 401, found [M + H] ¹H-NMR (DMSO- D₆): δ 1.046-1.068 (2H, m), 1.173-1.215 (2H, m), 3.401 (3H, s), 3.822- 3.856 (1H, m),7.380 (1H, dd, J = 8.1, 1.2 Hz), 7.450 (1H, d, J = 8.1 Hz), 8.107 (1H,d, J = 8.1 Hz), 8.363 (1H, q, J = 8.4 Hz) ¹⁹F (DMSO- D₆): δ −70.13,−70.03 71

9-cyclopropyl- 6-fluoro- 7-(3-hydroxy- phenyl)-8- methoxy-isothiazolo[5,4- b]quinoline- 3,4(2H,9H)- dione LCMS m/z calcd forC₂₀H₁₅FN₂O₄S ([M]⁺) 398; found M⁺ ¹H-NMR (DMSO- D₆): δ 1.032- 1.091 (m,2H), 1.162-1.203 (m, 2H), 3.372 (s, 3H), 3.808-3.842 (m, 1H), 6.856-6.935 (m, 3H), 7.331 (t, J = 7.8 Hz, 1H), 7.769 (d, J = 9.3 Hz, 1H),9.632 (brs, 1H) ¹⁹F (DMSO- D₆): δ −118.57 72

7-(4-aminophenyl)- 9- cyclopropyl- 6-fluoro-8- methoxy- isothiazolo[5,4-b]quinoline- 3,4(2H,9H)- dione LCMS m/z calcd for C₂₀H₁₆FN₃O₃S ([M]⁺)397, found M⁺ ¹H-NMR (DMSO- D₆): δ 1.042- 1.094 (2H, m), 1.165-1.206(2H, m), 3.375 (3H, s), 3.797- 3.868 (1H, m), 7.254 (2H, d, J = 8.1 Hz),7.514 (2H, d, J = 7.5 Hz), 7.783 (1H, d, J = 9.6 Hz); ¹⁹F: δ −118.97.¹⁹F (DMSO- D₆): δ −118.97 73

9-cyclopropyl- 6-fluoro- 8-methoxy- 7-(pyridin-3- yl)isothiazolo [5,4-b]quinoline- 3,4(2H,9H)- dione LCMS m/z calcd for C₁₉H₁₄FN₃O₃S ([M]⁺)383, found M⁺ ¹H-NMR (DMSO- D₆): δ 0.965- 1.017 (2H, m), 1.049-1.177(2H, m), 3.343 (3H, s), 3.739- 3.810 (1H, m), 7.728 (1H, dd, J = 2.7,7.8 Hz), 7.795 (1H, d, J = 9.3 Hz), 8.178 (1H, d, J = 8.1 Hz), 8.731(1H, d, J = 4.8 Hz), 8.819 (1H, brs) ¹⁹F (DMSO- D₆): δ −119.37

Example 10 Antimicrobial Activity of Compounds of the Invention

The antimicrobial activity of the compounds of the invention may beevaluated by a number of methods, including the following visual minimuminhibitory concentration (MIC) assay. This assay determines the minimumconcentration of compound required to inhibit growth of a bacterialstrain.

Minimum Inhibitory Concentration (MIC) Assay

Whole-cell antibacterial activity is determined by broth microdilutionusing conditions recommended by the NCCLS (see National Committee forClinical Laboratory Standards. 2001. Performance standards forantimicrobial susceptibility testing: 11^(th) informational supplement.Vol. 21, no. 1, M100-S11. National Committee for Clinical LaboratoryStandards, Wayne, Pa.). Test compounds are dissolved in DMSO and diluted1:50 in Mueller-Hinton II broth (Becton-Dickinson) to produce a 256μμg/ml stock solution. In a 96-well microtiter plate, the compoundsolution is serially two-fold diluted in Mueller-Hinton II broth. Afterthe compounds are diluted, a 50 μl aliquot of the test organism (˜1×10⁶cfu/mL) is added to each well of the microtiter plate. The final testconcentrations range from 0.125-128 μg/mL. Inoculated plates areincubated in ambient air at 37° C. for 18 to 24 hours. The organismsselected for testing included laboratory strains S. aureus ATCC 29213and E. coli ATCC 25922 (strains purchased from American Type CultureCollection, Manassas, Va.). The minimum inhibitory concentration (MIC)was determined as the lowest concentration of compound that inhibitedvisible growth of the test organism.

Certain compounds shown in Table I have an MIC of 1 μg/ml or lessagainst at least one of the S. aureaus and E. coli when tested in thisassay. Certain compounds disclosed in Table I exhibit an MIC of 100ng/ml or less against at least one of the S. aureaus and E. coli whentested in this assay.

Example 11 Cell Viability Staining with Alamar Blue

To determine whether the microcidal effect observed against S. aureusand E. coli is specific to bacterial cells, compounds are screened forcell viability effects on several human cell types.

Optimal cell density is first determined by plating cells in a 96-wellplate standard sterile tissue culture plates in 100 μl media, 10% FBS atsix cell densities from 500 cells/well to 15,000 cells/well. A cell freewell containing only media is used as a control. Cells are incubated at37° C. in a 5% CO2 incubator for 24 hours. 10% culture volume (10 ul) ofAlamar Blue (Biosource, DAL1100, 100 mL) is then added. Cells areincubated at 37° C. in a 5% CO₂ incubator and read in a Victor V platereader, 544 nm excitation, 590 nm emission, at 3, 4, and 24 hours afterthe addition of Alamar Blue. The cell number vs. change in fluorescenceis plotted to determine linearity of signal vs. cell number. The optimaldensity varies between 500-15,000 cells/well depending on the specificcell type. The optimal density is selected based on the highest numberof cells that is still in the linear response range.

Determination of Compound Cytotoxicity

Cells are plated at optimal cell density in a standard sterile tissueculture 96 well plate, and incubated at 37° C. O/N in a 5% CO₂incubator. 12 to 48 hours post-plating media is removed. The cells arewashed 1 or 2 times with 1×PBS and replaced with fresh media containingthe test compound in 1% DMSO. 24 to 72 hours after addition of compound,the media is removed, and the cells washed 1 to 2 times with 1×PBS.Fresh media containing 1/10 volume of Alamar Blue is then added. Platesare incubated 4 hours at 37° C. in a 5% CO₂ incubator and read in aVictor V plate reader, 544 nm excitation, 590 nm emission.

Compounds are diluted to 20 micromolar in 1% DMSO and media and screenedin duplicate to obtain single concentration cytotoxicity data. Eightconcentration points from 0.78 micromolar to 100 micromolar, run induplicate, are used to determine cyctotoxicity CC50 values. Cells with1% DMSO and media are used as a negative control, compounds having aknown CC50 against a particular cell type are used as positive controls.

The change in fluorescence vs. concentration of test compound is plottedto determine the cytotoxicity of the compound.

Sample media conditions, optimal plating densities, and positive controlcompounds for two cell types screened are presented in Table II.

Certain compounds disclosed in Example 1 to 6 and Example 9 exhibit CC50values greater than 10 uM against each of the cell lines listed belowwhen tested in this assay. Other cell types that may be used include butare not limited to Balb/3TC, CEM-SS, HeLa, HepG2, HT-29, MRC-5, SK-N-SH,U-87 MG, 293T, and Huh-7.

TABLE II Plating Positive Cell Line Media Density Control CHO 1. F-12Nutrient Mixture (Gibco 7,000 Terfenadine (Chinese #11765-054)containing cells/well CC₅₀ = hamster 10% FBS, 1% Pen Strep, 4.3-6.5 μMovary) 1.5 g/L Sodium Bicarbonate 2. McCoy's 5a medium, 10% FBS andPS/Gln Hep 2 Minimum Essential Medium - 7,000 Terfenadine (laryngealAlpha Medium (Gibco cells/well CC₅₀ = carcinoma) # 12571-063) containing3-5 μM 10% FBS, 1% Pen Strep, 1.5 g/L Sodium Bicarbonate

Example 12 Pharmaceutical Formulations

Examples 12A through 12G are examples of pharmaceutical compositionscontaining the compounds of Formula I. The abbreviation ‘A.M.’ standsfor an antimicrobial compound of the present invention.

Example 12A Oral Drops

5 grams of A.M. is dissolved in 5 ml of 2-hydroxypropanoic acid and 15ml polyethylene glycol at about 60°-80° C. After cooling to about30°-40° C., 350 ml polyethylene glycol is added and the mixture wasstirred well. A solution of 17.5 g sodium saccharin in 25 ml purifiedwater is then added. Flavor and polyethylene glycol q.s. (quantitysufficient) to a volume of 500 ml are added while stirring to provide anoral drop solution comprising 10 mg/ml of A.M.

Example 12B Capsules

20 grams of the A.M., 6 grams sodium lauryl sulfate, 56 grams starch, 56grams lactose, 0.8 grams colloidal silicon dioxide, and 1.2 gramsmagnesium stearate are vigorously stirred together. The resultingmixture is subsequently filled into 1000 suitable hardened gelatincapsules, comprising each 20 mg of the active ingredient.

Example 12C Film-Coated Tablets

Preparation of tablet core: A mixture of 10 grams of the A.M., 57 gramslactose and 20 grams starch is mixed well and thereafter humidified witha solution of 0.5 grams sodium dodecyl sulfate, and 1.0 gramspolyvinylpyrrolidone (KOLLIDON-K 90) in about 20 ml of water. The wetpowder mixture is sieved, dried, and sieved again. Then 100 gramsmicrocrystalline cellulose (AVICEL) and 15 grams hydrogenated vegetableoil (STEROTEX) are added. The whole is mixed well and compressed intotablets, giving 1000 tablets, each containing 10 mg of the activeingredient.

Coating: Ethyl cellulose (0.5 grams, ETHOCEL 22 CPS) in 15 ml ofdichloromethane is added to a solution of 1.0 grams methyl cellulose(Methocel 60 HG™) in 7.5 ml of denatured ethanol. Then 7.5 ml ofdichloromethane and 0.25 ml 1,2,3-propanetriol are added. Polyethyleneglycol (1.0 grams) is melted and dissolved in 7.5 ml of dichloromethaneand added to the cellulose-containing solution. Magnesium Octadecanoate(.25 grams), 0.5 grams polyvinylpyrrolidone, and 3.0 ml of concentratedcolor suspension (OPASPRAY K-1-2109) are added and the whole mixturehomogenized. The tablet cores are coated with this mixture in a coatingapparatus.

Example 12D Injectible Solutions

(i) 1.8 grams methyl 4-hydroxybenzoate and 0.2 grams propyl4-hydroxybenzoate are dissolved in about 0.5 L of boiling water. Aftercooling to about 50° C., 4 grams lactic acid, 0.05 grams propyleneglycol, and 4 grams of the A.M are added while stirring. The solution iscooled to room temperature and supplemented with water for injectionq.s. giving a solution containing 4 mg/ml of A.M. The solution issterilized by filtration and filled in sterile containers.

(ii) 100.0 g of an acid salt of an A.M. of the invention is dissolved inboiling water. After cooling to about 50° C., 37.5 grams lactic acid(90% by weight) are added while stirring. The solution is cooled to roomtemperature and water is added to 1 L. The solution is sterilized byfiltration and filled in sterile containers.

(iii) 5.00 g of an acid salt of an A.M. of the invention is dissolved inboiling water. After cooling to about 50° C., 2.20 grams lactic acid(90% by weight) are added while stirring. The solution is cooled to roomtemperature and water is added to 100 ml.

Example 12E Gel

A compound or salt of the invention may be formed as a gel for topicalapplication.

A gel is prepared by suspending A.M (0.2 g-5.0 g) in benzyl alcohol atroom temperature. A mixture of hydroxypropyl cellulose (2.5) grams anddemineralized water (q.s. 100 g) is added to the suspension withstirring.

Example 12F Cream

Phase I contains Sorbitan monostearate (2.0 g), Polyoxyethylene (20)sorbitan monostearate (1.5 g), Synthetic spermaceti (3.0 g) Cetylstearyl alcohol (10.0 g) and 2-Octyldodecanol (13.5 g). The phase Imixture is heated to 75° C., stirred and mixed.

Phase II contains A.M. (1.0 g). Phase II is added to phase I, stirredand suspended.

Phase III contains Benzyl alcohol (1.0 g) and demineralized water (q.s.100 g). Phase III is heated to 75° C. and added to phase II. The creamis mixed intensively and cooled slowly to room temperature, with furtherstirring. After cooling to room temperature the cream is homogenized.

Example 12G Sprays

The active compound solutions or suspensions prepared according toExample 12D can also be processed to sprays. For this purpose, forexample, a 60 to 90% active compound solution is mixed with 20 to 40% ofthe usual propellants, for example N₂, N₂O, CO₂, propane, butane,halogenohydrocarbons and the like.

1. A compound or pharmaceutically acceptable salt thereof, wherein thecompound is:9-cyclopropyl-6-fluoro-8-methoxy-7-(2-methylpyridin-4-yl)isothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;9-cyclopropyl-6-fluoro-8-methoxy-7-(6-methylpyridin-3-yl)isothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;9-cyclopropyl-6-fluoro-8-methoxy-7-((R)-1-methylisoindolin-5-yl)isothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;7-(3-amino-4-fluorophenyl)-9-cyclopropyl-8-methoxyisothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;9-cyclopropyl-8-methoxy-7-(2-methylpyridin-4-yl)isothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;(E)-4-(9-cyclopropyl-6-fluoro-3,4-dioxo-2,3,4,9-tetrahydroisothiazolo[5,4-b]quinolin-7-yl)picolinaldehydeO-methyl oxime;9-cyclopropyl-6-fluoro-7-(2-(hydroxymethyl)pyridin-4-yl)isothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;9-cyclopropyl-6-fluoro-7-(6-fluoropyridin-3-yl)-8-methoxyisothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;9-cyclopropyl-6-fluoro-7-(2-fluoropyridin-3-yl)-8-methoxyisothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;9-cyclopropyl-6-fluoro-7-(2-fluoropyridin-4-yl)-8-methoxyisothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;9-cyclopropyl-7-(4-(hydroxymethyl)phenyl)-8-methoxyisothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;9-cyclopropyl-7-(3-hydroxyphenyl)-8-methoxyisothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;7-(4-aminophenyl)-9-cyclopropyl-8-methoxyisothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;7-(4-amino-3,5-dimethylphenyl)-9-cyclopropyl-6-fluoro-8-methoxyisothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;7-(4-amino-3,5-dimethylphenyl)-9-cyclopropyl-6-fluoroisothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;9-cyclopropyl-6-fluoro-7-(4-(hydroxymethyl)phenyl)-8-methoxyisothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;9-cyclopropyl-7-(2-fluoropyridin-4-yl)-8-methoxyisothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;9-cyclopropyl-6-fluoro-7-(2-methylpyridin-4-yl)isothiazolo[5,4-b][1,8]naphthyridine-3,4(2H,9H)-dione;9-cyclopropyl-7-(2,6-dimethylpyridin-4-yl)-6-fluoroisothiazolo[5,4-b][1,8]naphthyridine-3,4(2H,9H)-dione;9-cyclopropyl-8-methoxy-7-(pyridin-3-yl)isothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;5-(9-cyclopropyl-6-fluoro-3,4-dioxo-2,3,4,9-tetrahydroisothiazolo[5,4-b]quinolin-7-yl)nicotinonitrile;5-(9-cyclopropyl-8-methoxy-3,4-dioxo-2,3,4,9-tetrahydroisothiazolo[5,4-b]quinolin-7-yl)nicotinonitrile;5-(9-cyclopropyl-6-fluoro-8-methoxy-3,4-dioxo-2,3,4,9-tetrahydroisothiazolo[5,4-b]quinolin-7-yl)nicotinonitrile;9-cyclopropyl-7-(6-fluoro-2-methylpyridin-3-yl)-8-methoxyisothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;9-cyclopropyl-6-fluoro-7-(6-fluoro-2-methylpyridin-3-yl)isothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;9-cyclopropyl-7-(2,6-dimethylpyridin-3-yl)-8-methoxyisothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;9-cyclopropyl-6-fluoro-7-(pyridin-4-yl)isothiazolo[5,4-b][1,8]naphthyridine-3,4(2H,9H)-dione;9-cyclopropyl-6-fluoro-7-(6-methylpyridin-3-yl)isothiazolo[5,4-b][1,8]naphthyridine-3,4(2H,9H)-dione;9-cyclopropyl-6-fluoro-7-(pyridin-3-yl)isothiazolo[5,4-b][1,8]naphthyridine-3,4(2H,9H)-dione;7-(4-(aminomethyl)phenyl)-9-cyclopropyl-6-fluoroisothiazolo[5,4-b][1,8]naphthyridine-3,4(2H,9H)-dione;9-cyclopropyl-7-(2,6-difluoropyridin-3-yl)-8-methoxyisothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;9-cyclopropyl-6-fluoro-7-(3-hydroxyphenyl)-8-methoxyisothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;7-(4-aminophenyl)-9-cyclopropyl-6-fluoro-8-methoxyisothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione;or9-cyclopropyl-6-fluoro-8-methoxy-7-(pyridin-3-yl)isothiazolo[5,4-b]quinoline-3,4(2H,9H)-dione.2. An anti-bacterial composition comprising a compound or salt of claim1 together with a carrier, diluent, or excipient.
 3. A pharmaceuticalcomposition comprising a compound or salt of claim 1 together with apharmaceutically acceptable carrier, diluent, or excipient.
 4. Apharmaceutical composition of claim 3, wherein the composition isformulated as an injectable fluid, an aerosol, a cream, a gel, a pill, acapsule, a tablet, a syrup, a transdermal patch, or an ophthalmicsolution.
 5. A composition comprising a compound or salt of claim 1 incombination with another one or more other antibacterial agent,antiprotozoal agent, antifungal agent, antiviral agent, interferon,efflux-pump inhibitor, or beta-lactamase inhibitor.
 6. A packagecomprising the pharmaceutical composition of claim 3 in a container andinstructions for using the composition to treat a patient suffering froma microorganism infection.
 7. The package of claim 6 wherein theinstructions are instructions for using the composition to treat apatient suffering from a bacterial infection.
 8. A method for treating abacterial infection in an animal comprising administering to the animala therapeutically effective amount of a compound or salt of claim
 1. 9.The method of claim 8 wherein the bacterial or prtozoal infection is aurinary tract infection, pyelonephritis, lower respiratory tractinfection, skin infection, skin-structure infection, urethral gonococcalinfection, cervical gonococcal infection, urethral chlamydial infection,cervical chlamydial infection, bone infection, joint infection,gram-negative bacterial infection, infectious diarrhea, typhoid fever,prostatitis, acute sinusitis, acute exacerbation of chronic bronchitis,pneumonia, intra-abdominal infection, gynecologic infection, or pelvicinfection.
 10. The method of claim 8 wherein the animal is a fish,amphibian, reptile, bird, or mammal.
 11. The method of claim 8 whereinthe animal is a mammal.
 12. The method of claim 11 wherein the mammal isa human.